Phosphocholine derivative inhibitors of phospholipase A2

ABSTRACT

Phospocholine derivatives having the formula: ##STR1## in which W, Z, Q and R are described in the specification are disclosed as useful for inhibiting the enzyme phospholipase A 2 . Methods of making and using the compounds are also disclosed.

This is a divisional of co-pending application Ser. No. 07/612,745 filedon Nov. 13, 1990 now U.S. Pat. No. 5,144,045.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The invention relates to novel phosphocholine derivatives which areinhibitors of the enzyme phospholipase A₂.

2. Description of the Prior Art

The enzyme phospholipase A₂ (phosphalide 2-acylhydrolase, PCA₂) isgenerally accepted as being the key enzyme responsible for exertingrate-determining regulatory control over the release of polyunsaturatedfatty acids from phospholipids [Flower, R. Q., et al., Biochem.Pharmac., 25, 285 (1976); Lapetina, E. G., et al., Annu. Rep. Med.Chem., 19, 213 (1984)]. This enzyme has been isolated from many sourcesand is known to cleave snglycerophospholipids at the 2-position thusgiving rise ultimately to the biosynthesis of platelet activatingfactor, prostaglandins, leukotrienes and prostacyclin [Dennis, E. A.,The Enzymes; Academic Press: New York; Vol. 16, Chapter 9 (1983)]. Theroles of these mediators in the inflammatory response associated withvarious diseases and physiological states is well known. Included amongthese diseases are asthma, arthritis, pancreatitis, myocardial ischemia,inflammation, pain, edema and their inflammatory related disorders.

Various compounds which are known to be inhibitors of phospholipase A₂have been described in U.S. Pat. No. 4,959,357, 4,933,365 and inWilkenson, Anti Inflammatory Phospholipase A₂ Inhibitors, Drugs of TheFuture, Vol. 15, No. 2, p 139-148 (1990). Various phosphocholinederivative compounds having anti hypertension activity have beendisclosed in U.S. Pat. Nos. 4,640,913, 4,697,031, 4,699,990, 4,703,130,4,939,127, 4,762,942, 4,883,816 and 4,894,367.

BRIEF SUMMARY OF THE INVENTION

The invention comprises novel phospholipase A₂ inhibiting compounds ofthe formula: ##STR2## wherein: A) n is an integer from 2 to 6;

B) R is selected from the group consisting of C₁ -C₄ alkyl;

C) Q is ##STR3## wherein R' is selected from the group consisting of C₁-C₂₄ alkyl, and W is a radical selected from the group consisting ofhydrogen and hydroxyl and Z is CH₂ --O--X wherein X is selected from thegroup consisting of C₁ -C₂₄ alkyl;

D) Q is selected from the group consisting of ##STR4## and-CH₂ --S--R'wherein R' is selected from the group consisting of C₁ -C₂₄ alkyl and Wis a radical selected from the group consisting of hydrogen and hydroxyland Z is selected from the group consisting of hydrogen and CH₂ --OXwherein X is selected from the group consisting of C₁ -C₂₄ alkyl;

E) Q is selected from the group consisting of:

(i) ##STR5## wherein m is O or 1, A and A' may or may not be the samebut are each selected from the group consisting of C₁ -C₂₄ alkyl,phenyl, C₁ -C₅ alkoxy phenyl, C₁ -C₅ alkyl phenyl, halogen substitutedphenyl and triflouromethylphenyl; ##STR6## wherein M is O or 1, A isselected fron the group consisting of C₁ -C₂₄ alkyl, phenyl, C₁ -C₅alkoxy phenyl, C₁ -C₅ alkyl phenyl, halogen substituted phenyl andtrifluoromethylphenyl;

(iii) --C.tbd.C--R' wherein R' is selected from the group consisting ofC₁ -C₂₄ alkyl;

(iv) ##STR7## wherein R' is selected from the group consisting of C₁-C₂₄ alkyl;

(v) --CH₂ --B(OH)₂

(vi) --CH₂ --C═CH₂

(vii) ##STR8## (viii) ##STR9## (ix) ##STR10## wherein R' is selectedfrom the group consisting of C₁ -C₂₄ alkyl;

(x) ##STR11## wherein R' is selected from the group consisting of C₁-C₂₄ alkyl;

(xi) ##STR12## wherein R' is selected from the group consisting of C₁-C₂₄ alkyl and J is selected from the group consisting of chlorine,bromine, fluorine or hydrogen;

(xii) ##STR13## wherein R' is selected from the group consisting of C₁-C₂₄ alkyl;

(xiii) ##STR14## wherein R' is selected from the group consisting of C₁-C₂₄ alkyl;

(xiv) ##STR15## wherein R' is selected from the group consisting of C₁-C₂₄ alkyl;

and W is the radical hydrogen and Z is--CH₂ --O--X wherein X is selectedfrom the group consisting of C₁ -C₂₄ alkyl;

F) W and Q are contained within a β-lactam ring: ##STR16## and Z isselected from the group consisting of C₁ -C₂₄ alkyl, G) W and Q arecontained within an isoazaline ring: ##STR17## and Z is --CH₂ --O--Xwherein X and R' are selected from the group consisting of C₁ -C₂₄alkyl.

The invention also comprises novel methods of makeing the phospholipaseA₂ inhibiting compounds described hereinabove.

The novel compounds described hereinabove have been proven to bespecific inhibitors of the enzyme phospholipase A₂. These compounds aretherefore useful for the treatment of any disease state or physiologicaldisorder that results from the direct action of phospholipase A₂ or themediators produced as a result of its activity. Consequently, use ofthese compounds for the treatment of pancreatitis, myocardial ischemia,inflammation, pain, edema, asthma, arthritis and any other disorderrelated to inflammation is indicated. The invention thus also comprisessuch methods of treatment.

DETAILED DESCRIPTION OF THE INVENTION

The preparation of the phospholipase A₂ inhibiting compounds accordingto the present invention is described hereinbelow in Flowcharts Athrough M wherein n, m, X, R', R', A, A' and J are as describedhereinabove.

As indicated in the sequence of reactions outlined in Flowchart A, thealcohol 2 is reacted with acid 3 in the presence of an agent such asdicyclohexylcarbodiimide with a base such as N,N-dimethylaminopyridinein an inert solvent such as dichloromethane to produce ester 5.Alternatively, alcohol 2 can be reacted with anhydride 4 in the presenceof a base such as sodium acetate in an inert solvent such as chloroformto produce ester 5. ##STR18##

As indicated in the sequence of reactions outlined in Flowchart B, andα,β unsaturated ester like ethyl acrylate is reacted with thiol 7 in thepresence of a base such as triethylamine in a solvent such as ethylalcohol to produce the sulfide-ester 8. When 8 is reacted with areducing agent such as lithium borohydride in a solvent such astetrahydrofuran, the alcohol-sulfide 9 is produced. ##STR19##

The alcohol-sulfide 9 is reacted with 2-bromoethyl phosphorodichlorodatein an inert solvent such as carbon tetrachloride in the presence of abase such as triethylamine. After approximately ten hours, the solventis evaporated and a polar aprotic solvent such as tetrahydrofuran isadded followed by an approximately equal volume of 0.5M sodium acetate.After approximately 10 hours, the solution is acidified with an acidsuch as hydrochloric acid and then extracted with an inert organicsolvent such as ether. The residue which remains after evaporation ofthe organic solvent is dissolved in a solvent mixture such aschloroform, 2-propanol, dimethylformamide and trimethylamine is added tothis mixture. After heating for approximately 5 hours at approximately60° C., the phosphocholine derivative 10 is produced [Wissner, A., etal., J. Med. Chem., 27. 1174 (1984)]. When compound 10 is reacted withone equivalent of an oxidizing agent such as 3-chloroperoxybenzoic acidin an inert solvent such as dichloromethane at a reduced temperature ofapproximately -78° C., the sulfoxide 11 is produced. When the sulfide 10is reacted with an excess amount of an oxidant such as ozone, whereinthe intermediate ozonide must be reduced with a reductant such asdimethylsulfide, at a reduced temperature of approximately -78° C. in aninert solvent such as dichloromethane, the sulphone 12 is produced.##STR20##

As indicated in the sequence of reactions outlined in Flowchart C, thefunctionalized ester 13 [previously described: J. Organometal. Chem.308(3), C₂₇ -C₃₂ (1986); Aust. J. Chem. 34(11), 2355 (1981); Pat. Specf.(Aust.) AU534112 B2, Jan. 5, 1984, 22 pp.] is reacted with alcohol 14 inthe presence of a base such as N,N-dimethylaminopyridine in an inertsolvent such as dichloromethane to produce 15. Compound 15 is reactedwith thiol 7 in the presence of a base such as triethylamine in asolvent such as ethyl alcohol to produce 16. When compound 16 is reactedwith a reducing agent such as lithium borohydride in a solvent such astetrahydrofuran, alcohol 17 is produced. Following the sequence ofreactions previously described in connection with Flowchart B, thealcohol 17 is converted to phosphocholine derivative 18. Reaction ofsulfide 18 with one equivalent of an oxidizing agent such as3-chloroperoxybenzoic acid in an inert solvent such as dichloromethaneat a reduced temperature of approximately -78° C. produces sulfoxide 19.Reaction of sulfide 18 with an excess amount of an oxidizing agent suchas 3-chloroperoxybenzoic acid in an inert solvent such asdichloromethane at a reduced temperature of approximately -78° C.,produces sulphone 20. ##STR21##

As indicated in the sequence of reactions outlined in Flowchart D,wherein L is selected from the group consisting ofp-methoxytriphenylmethyl or benzyl, the alcohol 21 is reacted withphosphate 22 in the presence of a base such asN-methyl-5-chloroimidazole in a solvent system such asacetonitrile/dichloromethane to produce 23. Upon removal of the group Lfrom 23 using, for example, hydrogenolytic conditions whereby compound23 is placed under an atmosphere of hydrogen in the presence of acatalyst such as palladium on carbon in a solvent system such ascyclohexane/acetic acid; the alcohol 24 is produced. When 24 issubjected to the same set of reactions described hereinabove inconnection with Flowchart B for attachment of a phosphocholine moiety,the phosphocholine derivative 25 is produced.

Alternatively, as indicated in the sequence of reactions outlined inFlowchart E the monomethyl derivative 29 may be prepared wherein X, A,L, m and n are as described hereinabove. This compound is useful forproviding a route to the mono-acid 30. ##STR22##

As indicated in the sequence of reactions outlined in Flowchart E, thealcohol 21 is reacted with dichlorophosphate 26 in the presence of abase such as dimethylaminopyridine and another base such astriethylamine in an inert solvent such as carbon tetrachloride. Thismixture is then reacted with methyl alcohol to produce 27. Upon removalof the group L using, for example, hydrogenolytic conditions wherebycompound 27 is placed under an atmosphere of hydrogen in the presence ofa catalyst such as palladium on carbon in a solvent system such ascyclohexane, acetic acid, the alcohol 28 is produced. When 28 issubjected to the sequence of reactions described hereinabove inconnection with Flowchart B for attachment of a phosphocholine moiety,compound 29 is produced. Reaction of 29 with an excess amount oftrimethylamine at an elevated temperature produces 30.

Referring to Flowchart F, X is as described hereinabove as C₁ -C₂₄ alkyland Y is a leaving group selected from the group consisting of chlorine,bromine, iodo, OSO₂ CH₃, and OSO₂ -4-methylphenyl. ##STR23##

As indicated in the sequence of reactions outlined in Flowchart F, thealcohol 31 [Corey, E. J., et al., Tett. Lett., 3775 (1975)] is reactedwith X-Y 32 wherein X is C₁ -C₂₄ alkyl and Y is a leaving group selectedfrom the group consisting of chlorine, bromine, iodo, OSO₂ CH₂ and OSO₂-4-methylphenyl in a solvent such as dimethylformamide in the presenceof a base such as sodiumhydride to produce 33. Alcohol 33 is subjectedto the same sequence of reactions described hereinabove in connectionwith Flowchart B for attachment of a phosphocholine moiety to produce34. When the olefinic moiety of 34 is reacted with an oxidant such asdiborane wherein the intermediate borohydride is hydroolyzed with water,boronic acid 35 is produced. ##STR24##

As indicated in the sequence of reactions outlined in Flowchart G, themalonate derivative 36 is reacted with a reductant such as lithiumaluminum hydride in a solvent such as diethyl ether to produce the diol37. When 37 is reacted with X-Y 32 in the presence of a base such assodium hydride in a solvent mixture such as dimethylformamide(DMF)-tetrahydrofuran (THF), alcohol 38 is produced. When 38 issubjected to the sequence of reactions which have been previouslyoutlined hereinabove in connection with Flowchart B for attachment of aphosphocholine group, 39 is produced. Reaction of 39 with an oxidantsuch as m-chloroperoxybenzoic acid in an inert solvent such asdichloromethane produces epoxide 40; whereas reaction of 39 with anoxidant such as ozone in a solvent system such as methylalcohol-dichloromethane and subsequent reduction of the intermediateozonide, inherent to such as oxidant, with a reductant such astrimethylphosphite produces 41. Reaction of 41 with hydroxylaminehydrochloride in the presence of a base such as sodium carbonate in asolvent system such as ethyl alcohol-water produces 42. ##STR25##

As indicated in the sequence of reactions in Flowchart H, alcohol 43 isreacted with X-Y 32 in the presence of a base such as sodium hydride andin a solvent system such as dimethylformamide (DMF)-tetrahydrofuran(THF) to produce 44. Acetal 44 is converted to an alcohol via reactionwith a reductant such as lithium aluminum hydride boron trifluorideetherate to produce 45. The hydroxyl group of 45 is functionalized tobecome a leaving group with an agent such as p-toluenesulfonylchloridein the presence of a base such as pyridine to produce 46. When 46 isreacted with sodium azide in a solvent such as dimethylformamide (DMF),the azide derivative 47 is formed. Reduction of the azide group of 47with a reductant such as lithium aluminum hydride in a solvent such asdiethylether produces the amine 48. Reaction of 48 with anhydride 49 inthe presence of a base such as dimethylaminopyridine in an inert solventsuch as chloroform produces 50. Removal of the benzyl group viareduction using, for example, hydrogen and palladium on carbon in asolvent system such as cyclohexane-acetic acid, produces 51. When 51 issubjected to the sequence of reactions described hereinabove inconnection with Flowchart B for attachment of a phosphocholine moiety,52 is produced. ##STR26##

As indicated in the sequence of reactions in Flowchart I, alcohol 53 isreacted with acid 54 in the presence of a base such as sodium hydride inan inert solvent such as toluene. This reaction mixture is thenhydrolyzed with an acidic methanol solution to produce 55. When 55 issubjected to the sequence of reactions previously described hereinabovein connection with Flowchart B for attachment of a phosphocholine moietyand this reaction mixture is treated with compound 56 (R'--OH wherein R'is selected from the group consisting of C₁ -C₂₄ alkyl), the ester 57 isproduced.

Referring to Flowchart J, R', X and Y are as described hereinabove and Mis selected from the group consisting of alkyl C₁ -C₅. ##STR27##

As indicated in the sequence of reactions in Flowchart J, malonatederivative 58 is reacted With a base such as sodium hydride in an inertsolvent such as tetrahydrofuran (THF). This reaction mixture is thenreacted with a reductant such as lithium aluminum hydride to produce 59.The hydroxyl group of 59 is converted to a leaving group such as bromidevia reaction with an agent such as phosphorous tribromide to produce 60.When 60 is reacted with a salt of 58 such as the sodium salt 58a in asolvent such as tetrahydrofuran (THF), 61 is produced. Reaction of 61with a reductant such as lithium aluminum hydride in an inert solventsuch as diethylether, produces diol 62. Deprotection of 62 with a basesuch as sodium hydride in a solvent such as dimethylformamide with X-Y32 being present, produces 63. When alcohol 63 is subjected to thesequence of reactions previously described hereinabove in connectionwith Flowchart B for transformation of a hydroxyl group to aphosphocholine, 64 is produced. Reaction of 64 with an oxidant such asozone wherein the intermediate ozonide is reduced with dimethylsulfideproduces ketone 65 or by reaction with m-chloroperbenzoic acid givesepoxide 64a. When 65 is subjected to reductive amination conditions suchas ammonium acetate sodium cyanoborohydride, the amine 66 is produced.When 65 is treated with borane, alcohol 65a is formed. Treatment of 65with methylmagnesium bromide affords alcohol 65b.

Referring to Flowchart K, R', X, Y and M are as described hereinabove.##STR28##

As indicated in the sequence of reactions outlined in Flowchart K,propargyl alcohol is reacted with a base such as lithium amide and thenwith R'-Y 67 to produce 68. The hydroxyl moiety of 68 is transformed toa leaving group such as bromide via the action of a reagent such asphosphorous tribromide in an inert solvent such as diethylether to give69. Reaction of the propargylic bromide derivative 69 with 58a in asolvent such as ethyl alcohol produces 70. Reduction of the carboxylategroups of 70 with a reductant such as lithium aluminum hydride in aninert solvent such as diethylether produces 71. When 71 is treated witha base such as sodium hydride in a solvent such as dimethylformamide(DMF) and then reacted with X-Y 32, compound 72 is produced. Subjectionof the alcohol 72 to the sequence of reactions previously described inconnection with Flowchart B for transformation of an alcohol moiety to aphosphocholine moiety prduces 73. Reaction of 73 with potassiumpermanganate affords dione 73a. ##STR29##

As indicated in sequence of reactions outlined in Flowchart L, the diol31 [Corey, E. J., et al, Tett. Lett., 3775 (1975)] is reacted with abase such as sodium hydride in a solvent such as dimethylformamide inthe presence of X-Y 32 to produce 33. Protection of the hydroxyl groupof 33 with a moiety such as t-butyldimethylsilyl is accomplished byreaction of 33 with t-butyldimethylsilylchloride in a solvent such asdichloromethane in the presence of a base such as imidazole to produce74. Compound 74 is reacted with a nitrile oxide which can be generated,for example, from nitro compound 75 and a dehydrating agent such asphenyl isocyanate to produce 76. Removal of the t-butyldimethylsilylprotecting group with a fluoride source such as tetra-n-butyl-ammoniumfluoride in a solvent such as tetrahydrofuran (THF) produces 77. When 77is subjected to the sequence of reactions previously described inconnection with Flowchart B for attachment of a phosphocholine group,the phosphocholine derivative 78 is formed. Reduction of the isoxazolinering using a catalyst such as palladium on carbon under an atmosphere ofhydrogen in a solvent such as methanol produces 79. ##STR30##

As indicated in the sequence of reactions outlined in Flowchart M, thealcohol 59 is reacted with a base such as sodium hydride in a solventsuch as dimethylformamide (DMF) in the presence of benzyl bromide toproduce 80. The olefinic moiety of 80 is transformed into a β-lactamring via reaction with chlorosulfonylisocyanate and subsequent reductionof the intermediate with a reductant such as sodium sulfite to produce81. Removal of the benzyl group of 81 under hydrogenolytic conditionsusing a catalyst such as palladium-on-carbon produces 82. Reaction of 82with 2-chloro-2-oxo-1,3,2-dioxaphospholane [Edmundson, R. N.; Chem. Ind.(London) 1962, 1828] in the presence of a base such as triethylamineproduces a cyclic phosphate which is opened by reaction withtrimethylamine to produce 83.

The syntheses of the various starting materials cited hereinabove havebeen described previously and, in addition, require for their synthesisthe use of synthetic methods and procedures widely known to thoseskilled in the art. Furthermore, descriptions of various applicablepreparations have been described in U.S. Pat. Nos. 4,640,913, 4,697,031,4,703,130, 4,699,990, 4,762,942, 4,883,816 and 4,894,367, copendingapplication Serial No. 679,790 filed Dec. 10, 1984 and Tereshita, Z., etal., Life Sci., 32, 1975 (1983). The above patents, applications andcitations describe the preparations of both optically active and racemicmaterials, therefore, the compounds of this invention can be preparedboth as racemates and as individual R and S enantiomers.

The compounds of this invention were tested for pharmacological activityas described in the following tests.

IN VITRO PORCINE PANCREATIC PHOSPHOLIPASE (PLA2) ASSAY

Phospholipase A₂ activity was measured by a radiochemical assay in whichlabeled synthetic phosphatidylcholine substrate is converted by porcinepancreatic PLA₂ to free radiolabeled arachidonate and lyso PC.Radiolabeled arachidonate is extracted from the reaction mixture andseparated from unreacted substrate by column chromatography or TLC.Radioactivity (cpm) in samples is quantitated by liquid scintillationcounting. Specific details are as follows:

Labeled substrate(1-stearyl-2-³ H-arachidonylphosphatidylcholine;Amersham, sp. act. 84 (uCi/mmole, 0.025 to 0.2 uCi/assay tube andunlabeled substrates (Avanti), in sufficient quantity to give 20,000 cpmto 50,000 cpm and 10 μM to 1,000 μM final concentration/assay tube aredried under a stream of nitrogen. The PLA₂ and mPLA₂ assays utilized a 2nM substrate concentration. The mHPLA₂ and HPLA₂ assays (vida infra)utilized a 10 μM substrate concentration and the SmPLA₂ assay utilized asubstrate concentration ranging from 100 to 1000 μm. The substrate isthen redissolved in 0.1M tris-HCl buffer, pH 9.0 containing 10 mM CaCl₂and 10 mM Na taurocholate detergent. The mixture is sonicated in aKontes Sonifier to yield a homogeneous dispersion. The "mixed" micellarsubstrate is clear. An aliquot is reserved for a cpm count (totalsubstrate). The total reaction volume is 500 μL; siliconized glasswareor plasticware is used throughout. Enzyme (Sigma, sp. act.>690 U/mgprotein and preferably 900 U/mg protein) is diluted 1:750 or 1:1000 in0.1M tris-HCl buffer, pH 9.0 (without Ca⁺⁺ and detergent). Enzymedilution depends on substrate concentration and required enzymeconcentration.

The substrate in buffer is kept on ice until the reaction is started.The reaction is started by adding 10 μL of diluted enzyme solution to490 μL of micellar substrate solution. Samples are incubated in a 37° C.water bath for 15 minutes with gentle mixing. The reaction is terminatedby the addition of 100 μL of 1M fresh trichloroacetic acid (TCA)solution and chilling the tubes on ice for 30 minutes.

Samples (600 μL) are extracted with 3 ml CHCl₃ /MeOH (2:1) for thinlayer chromatography or in hexane/ether (9:1) for column chromatography.The phases are separated by gentle centrifugation at 700 rpm×5'. Theorganic phase is recovered.

THIN LAYER CHROMATOGRAPHY

Labeled arachidonate extracted with CHCl₃ /MeOH is separated on silicagel G TLC plates (Analtech). Twenty μL of a 2 mg/ml internal standardsolution of arachidonic acid (Sigma) is combined with 2-3 ml of theorganic phase. The entire mixture is dried under nitrogen andredissolved in 50 μL of CHCl₃ /MeOH (2:1). A 20 μL sample is counted in10 ml of Beckman Redi-solv HP and spotted on silica gel G plates. Thesolvent system used to develop the plate is CHCl₃ /MeOH/13.5N NH₃ /H₂ O(70:30:4:1). Plates are developed in a glass chamber for 1-11/2 hours,air dried, and the spots visualized in an iodine vapor chamber.Standards of phosphatidylcholine, lyso PC, and arachidonic acid (Sigma)are made as 2 mg/ml stocks in CHCl₃ /MeOH (2:1) and spotted on plates.Spots corresponding to arachidonate and phospholipid are scraped andcounted in 10 ml Beckman Redi-Solv HP scintillation fluid.

COLUMN CHROMATOGRAPHY

Samples are extracted with hexane/ether and the upper organic phaserecovered. An aliquot is applied to a Silicar CC₄ (Malincrodt) column.Columns are prepared using 1 ml of a 1:1 slurry of silicar CC₄ inhexane/ether in a Pasteur pipet (approx. 2 cm height). Columns arerinsed with 500 μL of hexane/ether before adding the sample (300-600 μL)to the column. Columns are eluted with a total of 6 ml of hexane/ether.The 6 ml sample is counted in 10 ml of Beckman Redi-Solv HPscintillation fluid.

DRUG INHIBITORS IN ENZYME ASSAYS

or the phospholipase A₂ assay, compounds are prepared as 1 mg/ml stocksolutions in MeOH and diluted appropriately in PBS. Ten ul aliquots ofthe respective working stock solutions are either preincubated withphospholipase A₂ enzyme in buffer in the absence of Ca⁺⁺ or detergentfor a specific time (enzyme preincubation design) or are blown downunder a N₂ stream with the substrate prior to addition of buffer (mixedmicellar design). Reactions are always begun by addition of enzyme(preincubated with compound or diluent or fresh) to substrate dispersionin buffer.

The assays are grouped as follows:

PLA₂ Assay: preincubation of drug+enzyme; substrate conc=2 nM

MPLA₂ Assay: mixed micelle design of drug+substrate connc=2 nM

HPLA₂ Assay: preincubation design; [s]=10 μm

MHPLA₂ Assay: mixed micelle design [s]=10 μm

SMPLA₂ Assay: mixed micelle design; [s]=100-100 μM

The results of these tests are displayed in the following five tables.

                                      TABLE I                                     __________________________________________________________________________    MHPLA.sub.2                                                                   Compound          Dose μ/ml                                                                        % Inh.                                                                            Avg.                                                                             IC.sub.50 μ/ml                                                                   Avg.                                                                             Subs. con.                            __________________________________________________________________________    2,5,7-Trioxa-6-phosphanonan-9-                                                                  10     6              10                                    aminium, 1-carboxy-3-[(hexadecyl-                                                               25    13  30          10                                    oxy)methyl]-6-hydroxy-N,N,N-tri-                                                                      46              10                                    methyl-, hydroxide, inner salt,                                               6-oxide                                                                       N-[3-(Hexadecyloxy)-2-hydroxy-                                                                  25    35              10                                    methyl)propyl]octadecanamide                                                  4-[(Hexadecyloxy)methyl]-7-                                                                     25    33              10                                    hydroxy-N,N,N-trimethyl-1,1-                                                  diphenoxy-2,6,8-trioxa-1,7-                                                   diphosphadecan-10-aminium, hy-                                                droxide, inner salt, 1,7-dioxide                                              3.5-Dioxa-9-thia-4-phosphapenta-                                                                25    STIM            10                                    cosan-1-aminium, 4-hydroxy-N,N,N-                                             trimethyl-, hydroxide, inner salt,                                            4-oxide                                                                       7-[(Hexadecylthio)methyl]-4-                                                                    10    STIM            10                                    hydroxy-N,N,N-trimethyl-3,5,9-                                                                  20    STIM            10                                    trioxa-4-phosphapentacosan-1-                                                                   25    81              10                                    aminium, hydroxide, inner salt,                                               4-oxide                                                                       7-[(Hexadecylsulfonyl)methyl]-4-                                                                25    19              10                                    hydroxy-N,N,N-trimethyl-3,5,9-                                                trioxa-4-phosphapentacosan-1-                                                 aminium, hydroxide, inner salt,                                               4-oxide                                                                       4-Hydroxy-N,N,N-trimethyl-3,5-                                                                  10     0     62.0     10                                    dioxa-9-thia-4-phosphapentacosan-                                                               20    19              10                                    1-aminium, hydroxide, inner                                                                     25    24              10                                    salt, 4,9-trioxide                                                            4-Hydroxy-N,N,N-trimethyl-3,5-                                                                  25    16                                                    dioxa-9-thia-4-phosphapentacosan-                                             1-aminium, hydroxide, inner                                                   salt, 4,9-dioxide                                                             9-Ethoxy-7-[(hexadecyloxy)methyl]-                                                              10    18     56.2                                           N,N,N-trimethyl-4-hydroxy-3,5,10-                                                               10    18     163.0                                          trioxa-4,9-diphosphadodecan-1-                                                                  20    44                                                    aminium, hydroxide, inner salt,                                                                 25    45                                                    4,9-dihydroxide         42                                                                            34                                                    4-Hydroxy-N,N,N-trimethyl-7-(2-                                                                 25    19     10                                             propanyl)-3,5,9-trioxa-4-phospha-                                             pentacosan-1-aminium, hydroxide,                                              inner salt, 4-oxide                                                           4-Hydroxy-7-[2-hydroxyimino)                                                                    10     0              10                                    ethyl]-N,N,N-trimethyl-3,5,9-                                                                   20     8              10                                    trioxa-4-phosphapentacosan-1-                                                                   25    28              10                                    aminium, hydroxide, inner                                                     salt, 4-oxide                                                                 7-(Boronomethyl)-4-hydroxy-                                                                     25     9              10                                    N,N,N-trimethyl-3,5,9-trioxa-                                                 4-phosphapentacosan-1-aminium,                                                hydroxide, inner salt, 4-oxide                                                3,5,8,10-Tetraoxa-4,9-diphospha-                                                                10     0              10                                    hexacosan-1-aminium, 7-[(hexa-                                                                  25    STIM                                                  decyloxy)methyl]-4-hydroxy-9-                                                 methoxy-N,N,N-trimethyl-, hy-                                                 droxide, inner salt, 4,9-dioxide                                              3,5,8,10-Tetraoxa-4,9-diphospha-                                                                10     0                                                    hexacosan-1-aminium, 7-[(hexa-                                                                  25     0              10                                    decyloxy)methyl]-4-hydroxy-9-                                                                   25    24  62          10                                    methoxy-N,N,N-trimethyl-, hy-                                                                   25    100             10                                    droxide, inner salt, 4,9-dioxide                                                                25    STIM            10                                    3,5,8,10-Tetraoxa-4,9-diphospha-                                                                10    STIM            10                                    hexacosan-1-aminium, 7-[(hexa-                                                                  25    STIM            10                                    decyloxy)methyl]-4,9-dihydroxy-                                                                 25    STIM            10                                    N,N,N-trimethyl-, hydroxide,                                                  4-(inner salt), 4,9-dioxide                                                   3,5,8,10-Tetraoxa-4,9-diphospha-                                                                10    STIM            10                                    hexacosan-1-aminium, 7-[(hexa-                                                                  25    STIM            10                                    decyloxy)methyl]-4,9-dihydroxy-                                                                 25    STIM            10                                    N,N,N-trimethyl-, hydroxide,                                                  4-(inner salt), 4,9-dioxide, (7S)                                             3,5,8,10-Tetraoxa-4,9-diphospha-                                                                25     0              10                                    hexacosan-1-aminium, 7-[ (hexa-                                               decyloxy)methyl]-4-hydroxy-9-                                                 methoxy-N,N,N-trimethyl-, hy-                                                 droxide, inner salt, 4,9-dioxide,                                             7(S)-, upper diastereomer                                                     3,5,8,10-Tetraoxa-4,9-diphospha-                                                                25     7              10                                    hexacosan-1-aminium, 7-[(hexa-                                                decyloxy)methyl]-4-hydroxy-9-                                                 methoxy-N,N,N-trimethyl-, hy-                                                 droxide, inner salt, 4,9-dioxide,                                             (7R)-, upper diastereomer                                                     3,5,8,10-Tetraoxa-4,9-diphospha-                                                                25     1              10                                    hexacosan-1-aminium, 7-[(hexa-                                                decyloxy)methyl]-4,9-dihydroxy-                                               N,N,N-trimethyl-, hydroxide, 4-                                               (inner salt), 4,9-dioxide                                                     3,5,8,10-Tetraoxa-4,9-diphos-                                                                   25     9              10                                    phahexacosan-1-aminium, 7-[(hexa-                                             decyloxy)methyl]-4-hydroxy-9-                                                 methoxy-N,N,N-trimethyl-, hy-                                                 droxide, inner salt, 4,9-oxide,                                               (7S)-, lower diastereomer                                                     7-(2-Nonadecynyl)-4-hydroxy-                                                                    25    16              10                                    N,N,N-trimethyl-3,5,9-trioxa-                                                 4-phosphapentacosan-1-aminium,                                                hydroxide, inner salt, 4-oxide                                                7-(2,3-Dioxononadecyl)-4-hydroxy-                                                               25     0              10                                    N,N,N-trimethyl-3,5,9-trioxa-4-                                               phosphapentacosan-1-aminium, hy-                                              droxide, inner salt, 4-oxide                                                  3,5,8,10-Tetraoxa-4,9-diphos-                                                                   25     0              10                                    phapentadecan-1-aminium, 4-                                                   hydroxy-9-methoxy-N,N,N-tri-                                                  methyl-7-[(pentyloxy)-methyl]-,                                               hydroxide, inner salt, 4,9-dioxide                                            3,5,8,10-Tetraoxa-4,9-diphos-                                                                   25    12              10                                    phapentadecan-1-aminium, 4,9-                                                 dihydroxy-N,N,N-trimethyl-7-                                                  [(pentyloxy)methyl]-, hydroxide,                                              4-(inner salt), 4,9-dioxide                                                   7-(2-Aminooctadecyl)-4-hydroxy-                                                                 10    93     1.4      10                                    N,N,N-trimethyl-3,5,9-trioxa-4-                                                                 10    59  70 5.2   4.3                                                                              10                                    phosphapentacosan-1-aminium,                                                                    10    59     6.4      10                                    hydroxide, inner salt, 4-oxide                                                                  20    97              10                                                      25    78              10                                                      25    76  77          10                                                      25    78              10                                    3,5,8,10-Tetraoxa-4,9-diphospha-                                                                10    STIM                                                  pentadecan-1-aminium, 7-[(hexa-                                                                 25    STIM                                                  decyloxy)methyl]-4,hydroxy-                                                                     25    STIM                                                  9-methoxy-N,N,N-trimethyl-,                                                   hydroxide, 4-(inner salt),                                                    4,9-dioxide, less polar isomer                                                3,5,8,10-Tetraoxa-4,9-diphospha-                                                                10     0                                                    pentadecan-1-aminium, 7-[(hexa-                                                                 25    STIM                                                  decyloxy)methyl]-4,hydroxy-                                                                     25    STIM                                                  9-methoxy-N,N,N-trimethyl-,                                                   hydroxide, 4-(inner salt),                                                    4,9-dioxide, more polar isomer                                                3,5,8,10-Tetraoxa-4,9-diphospha-                                                                10    STIM                                                  pentadecan-1-aminium, 7-[(hexa-                                                                 25    STIM                                                  decyloxy)methyl]-4,9-dihydroxy-                                                                 25    STIM                                                  N,N,N-trimethyl-, hydroxide,                                                  4-(inner salt), 4,9-dioxide,                                                  3,5,9-Trioxa-4-phosphapentacosan-                                                               25    11                                                    1-aminium, 7-(3,3-difluoro-2-                                                 methylenenonadecyl)-4-hydroxy-                                                N,N,N-trimethyl-, hydroxide,                                                  inner salt, 4-oxide                                                           3,5,9-Trioxa-4-phosphapentacosan-                                                               10     0              10                                    1-aminium, 7-(3,3-difluoro-2-                                                                   20     9              10                                    oxononadecyl)-4-hydroxy-N,N,N-                                                                  25     8              10                                    trimethyl-, hydroxide, inner                                                  salt, 4-oxide                                                                 3,5,9-Trioxa-4-phosphapentacosan-                                                               25    11              10                                    1-aminium, 7-(3,3-difluoro-2-                                                 hydroxynonadecyl)-4-hydroxy-                                                  N,N,N-trimethyl-, hydroxide,                                                  inner salt, 4-oxide                                                           (2R)-2-[[[(2-Hexadecyl-4-oxo-2-                                                                 10    STIM            10                                    azetidinyl)methoxy]hydroxyphos-                                                                 20     0              10                                    phinyl]oxy]-N,N,N-trimethyl-                                                                    25    26              10                                    ethanaminium, hydroxide,                                                      inner salt                                                                    3,5,9-Trioxa-4-phosphapentacosan-                                                               10    35     7.9      10                                    1-aminium, 7-[(bromoacetyl)oxy]-                                                                10    24  32 26.6  50 10                                    4-hydroxy-N,N,N-trimethyl-,                                                                     10    35     35.5     10                                    hydroxide, inner salt, 4-oxide                                                                  10    34     130.0    10                                                      25    78              10                                                      25    29              10                                                      25    54  50          10                                                      25    43              10                                                      25    52              10                                                      25    50              10                                                      25    44              10                                    7-[(Chloroacetyl)oxy]-4-hydroxy-                                                                10    27              10                                    N,N,N-trimethyl-3,5,9-trioxa-4-                                                                 25     3              10                                    phosphapentacosan-1-aminium,                                                                    25    35   3          10                                    hydroxide, inner salt, 4-oxide                                                                  25     0              10                                    10-Bromo-7-[(hexadecyloxy)methyl]-                                                              10     3   8          10                                    4-hydroxy-N,N,N-trimethyl-9-oxo-                                                                10    14              10                                    3,5,8-trioxa-4-phosphahexacosan-                                                                25     0              10                                    1-aminium, hydroxide, inner                                                                           20  10          10                                    salt, 4-oxide            0              10                                                            19              10                                                      50    37  137         10                                    7-[(Hexadecylsulfinyl)methyl]-4-                                                                10     3              10                                    hydroxy-N,N,N-triimethyl-3,5,9-                                                                 20    22              10                                    trioxa-4-phosphapentacosan-1-                                                                   25    68              10                                    aminium, hydroxide, inner salt,                                               4-oxide                                                                       __________________________________________________________________________

                                      TABLE II                                    __________________________________________________________________________    SMPLA.sub.2                                                                   Compound          Dose μ/ml                                                                        % Inh.                                                                            Avg.                                                                             IC.sub.50 μ/ml                                                                   Avg.                                                                             Subs. con.                            __________________________________________________________________________    7-(2-Aminooctadecyl)-4-hydroxy-                                                                 25    63  61 15.1  20 100                                   N,N,N-trimethyl-3,5,9-trioxa-4-                                                                 25    59     25.0     100                                   phosphapentacosan-1-aminium,                                                  hydroxide, inner salt, 4-oxide                                                3,5,8,10-Tetraoxa-4,9-diphos-                                                                   10    STIM            100                                   phapentadecan-1-aminium,                                                                        10    7               500                                   7-[(hexadecyloxy)methyl]-4,9-                                                                   10    22              1000                                  dihydroxy-N,N,N-trimethyl-,                                                                     25    STIM            100                                   hydroxide, 4-(inner salt),                                                                      25    29              500                                   4,9-dioxide       25    3               1000                                                    50    STIM            100                                                     50    STIM            500                                                     50    STIM            1000                                  10-Bromo-7-[(hexadecyloxy)methyl]-                                                              10    8               100                                   4-hydroxy-N,N,N-trimethyl-9-oxo-                                                                10    0               500                                   3,5,8-trioxa-4-phosphahexacosan-                                                                10    5               1000                                  1-aminium, hydroxide, inner                                                                     25    1               100                                   salt, 4-oxide     25    0               500                                                     25    10              1000                                                    50    18              100                                                     50    8               500                                                     50    6               1000                                  __________________________________________________________________________

                                      TABLE III                                   __________________________________________________________________________    PLA.sub.2                                                                     Compound          Dose μ/ml                                                                        % Inh.                                                                            Avg.                                                                             IC.sub.50 μ/ml                                                                   Avg.                                                                             Subs. con.                            __________________________________________________________________________    2,5,7-Trioxa-6-phosphanonan-9-                                                                  20    44              2                                     aminium, 1-carboxy-3-[(hexa-                                                  decyloxy)methyl]-6-hydroxy-                                                   N,N,N-trimethyl-, hydroxide,                                                  inner salt, 6-oxide                                                           N-[3-(Hexadecyloxy)-2-hydroxy-                                                                  20    24              2                                     methyl)propyl]octadecanamide                                                                    25    93              2                                     4-[(Hexadecyloxy)methyl]-7-                                                                     10    75     4.8      2                                     hydroxy-N,N,N-trimethyl-1,1-                                                                    20    85                                                    diphenoxy-2,6,8-trioxa-1,7-                                                   diphosphadecan-10-aminium,                                                    hydroxide, inner salt, 1,7-dioxide                                            3,5-Dioxa-9-thia-4-phosphapenta-                                                                10    54     6.2      2                                     cosan-1-aminium, 4-hydroxy-                                                                     10    39              2                                     N,N,N-trimethyl-, hydroxide,                                                                    10    43  4.5         2                                     inner salt, 4-oxide                                                                             10    47              2                                                       10    44              2                                                       20    64              2                                     7-[(Hexadecylthio)methyl]-4-                                                                    10    60  26          2                                     hydroxy-N,N,N-trimethyl-3,5,9-                                                                  10    23              2                                     trioxa-4-phosphapentacosan-1-                                                                   10    14              2                                     aminium, hydroxide, inner salt,                                                                 10    20              2                                     4-oxide           10    13              2                                                       20    61     3.5      2                                     2-[[(2-Bromoethoxy)hydroxyphos-                                                                 10    42  21          2                                     phinyl]oxy]ethyl hexadecyl                                                                      10     1                                                    phosphoric acid, methyl ester                                                 7-[(Hexadecylsulfonyl)methyl]-4-                                                                10     9  57 2.3      2                                     hydroxy-N,N,N-trimethyl-3,5,9-                                                                  10    86              2                                     trioxa-4-phosphapentacosan-1-                                                                   10    76              2                                     aminium, hydroxide, inner salt                                                                  20    82              2                                     4-oxide                                                                       3,9-Dihydroxy-N,N,N-trimethyl-                                                                  10     4  20          2                                     3,5,8,10-tetraoxa-4,9-diphospha-                                                                10    37              2                                     hexacosan-1-aminium, hydroxide,                                               4-(inner salt) 4,9-dioxide                                                    4,9-Dihydroxy-N,N,N-trimethyl-                                                                  10     8  40          2                                     3,5,8,10-tetraoxa-4,9-diphos-                                                                   10    32              2                                     phaoctacosan-1-aminium, hy-                                                   droxide, 4-(inner salt),                                                      4,9-dioxide                                                                   4-Hydroxy-N,N,N-trimethyl-3,5-                                                                  10    65  69 4.9      2                                     dioxa-9-thia-4-phosphapenta-                                                                    10    74              2                                     cosan-1-aminium, hydroxide,                                                                     20    88              2                                     inner salt, 4,9-trioxide                                                      4-Hydroxy-N,N,N-trimethyl-3,5-                                                                  10    59  61 6.2      2                                     dioxa-9-thia-4-phosphapentaco-                                                                  10    63              2                                     san-1-aminium, hydroxide, inner                                                                 20    82              2                                     salt, 4,9-dioxide                                                             9-Ethoxy-7-[(hexadecyloxy)methyl]-                                                              10    71  74 5.0      2                                     N,N,N-triimethyl-4-hydroxy-                                                                     10    77              2                                     3,5,10-trioxa-4,9-diphosphado-                                                                  20    81              2                                     decan-1-aminium, hydroxide, inner                                             salt, 4,9-dihydroxide                                                         4-Hydroxy-N,N,N-trimethyl-7-                                                                    10    75  80 1.6      2                                     (2-propanyl)-3,5,9-trioxa-4-                                                                    10    85              2                                     phosphapentacosan-1-aminium,                                                                    20    91              2                                     hydroxide, inner salt, 4-oxide                                                4-Hydroxy-N,N,N-trimethyl-7-                                                                    10    71  74 2.4      2                                     (oxiranylmethyl)-3,5,9-trioxa-                                                                  10    77              2                                     4-phosphapentacosan-1-aminium,                                                                  20    88              2                                     hydroxide, inner salt, 4-oxide                                                4-Hydroxy-N,N,N-trimethyl-7-                                                                    10    47  43 13.2     2                                     (2-oxoethyl)-3,5,9-trioxa-4-                                                                    10    39              2                                     phosphapentacosan-1-aminium,                                                                    20    62              2                                     hydroxide, inner salt, 4-oxide                                                4-Hydroxy-7-[2-(hydroxyimino)                                                                   10    56  53 5.3      2                                     ethyl]-N,N,N-trimethyl-3,5,9-                                                                   10    49              2                                     trioxa-4-phosphapentacosan-1-                                                                   20    81              2                                     aminium, hydroxide, inner salt,                                               4-oxide                                                                       7-(Boronomethyl)-4-hydroxy-N,N,N-                                                               10    57     5.6      2                                     trimethyl-3,5,9-trioxa-4-phospha-                                                               20    77              2                                     pentacosan-1-aminium, hydroxide,                                              inner salt, 4-oxide                                                           3,5,8,10-Tetraoxa-4,9-diphospha-                                                                10    61  68 6.0   5.4                                                                              2                                     hexacosan-1-aminium, 7-[(hexa-                                                                  10    70     4.5      2                                     decyloxy)methyl]-4-hydroxy-9-                                                                   10    73     5.8      2                                     methoxy-N,N,N-trimethyl-,                                                                       20    65                                                    hydroxide, inner salt, 4,9-dioxide                                                              25    71  77                                                                  25    83                                                                      50    73  80                                                                  50    88                                                    3,5,8,10-Tetraoxa-4,9-diphospha-                                                                10    72  30 3.2   3  2                                     hexacosan-1-aminium, 7-[(hexa-                                                                  10    78     2.8      2                                     decyloxy)methyl]-4-hydroxy-9-                                                                   25    72  75          2                                     methoxy-N,N,N-trimethyl-, hy-                                                                   25    85              2                                     droxide, inner salt, 4,9-dioxide                                                                50    68  77          2                                                       50    87              2                                     4-Hydroxy-N,N,N-trimethyl-7-(2-                                                                 10    59  62 3.8      2                                     methyleneoctadcyl)-3,5,9-trioxa-                                                                10    65              2                                     4-phosphapentacosan-1-aminium,                                                                  20    52              2                                     hydroxide, inner salt, 4-oxide                                                4-Hydroxy-N,N,N-trimethyl-7-(2-                                                                 10    59  60 5.8      2                                     oxooctadecyl)-3,5,9-trioxa-4-                                                                   10    61              2                                     phosphapentacosan-1-aminium,                                                                    20    62              2                                     hydroxide inner salt, 4-oxide                                                 7-[(2-Hexadecyloxiranyl)methyl]-                                                                10    78     1.8      2                                     4-hydroxy-N,N,N-trimethyl-3,5,9-                                                                20    75              2                                     trioxa-4-phosphapentacosan-1-                                                 aminium, hydroxide, inner salt,                                               4-oxide                                                                       4-Hydroxy-7-(2-hydroxy-2-methyl-                                                                10    65  64 5.9      2                                     octadecyl)-N,N,N-trimethyl-3,5,9-                                                               10    62              2                                     trioxa-4-phosphapentacosan-1-                                                                   20    66              2                                     aminium, hydroxide, inner salt,                                               4-oxide                                                                       4-Hydroxy-7-(2-hydroxyoctadecyl)-                                                               10    80     1.6      2                                     N,N,N-trimethyl-3,5,9-trioxa-4-                                                                 20    80              2                                     phosphapentacosan-1-aminium,                                                  hydroxide, inner salt, 4-oxide                                                4-Hydroxy-7-(2-hydroxyoctadecyl)-                                                               10    80     1.7      2                                     N,N,N-trimethyl-3,5,9-trioxa-4-                                                                 20    80              2                                     phosphapentacosan-1-aminium,                                                  hydroxide, inner salt, 4-oxide                                                isomer                                                                        3,5,8,10-Tetraoxa-4,9-diphospha-                                                                10    19  22          2                                     hexacosan-1-aminium, 7-[(hexa-                                                                  10     9              2                                     decyloxy)methyl]-4-hydroxy-9-                                                                   10    48              2                                     methoxy-N,N,N-trimethyl-,                                                                       10    14              2                                     hydroxide, inner salt, 4,9-                                                                     20    32  34 13.7     2                                     dioxide (7R)-, lower diastereomer                                                               20    14              2                                                       20    55              2                                     3,5,8,10-Tetraoxa-4,9-diphospha-                                                                10    14  17          2                                     hexacosan-1-aminium, 7-[(hexa-                                                                  10     2              2                                     decyloxy)methyl]-4,9-dihydroxy-                                                                 10    34              2                                     N,N,N-trimethyl-, hydroxide,                                                                    20    15  16          2                                     4-(inner salt), 4,9-dioxide, (7S)                                                               20     2              2                                                       20    32              2                                                       25     8              2                                     3,5,8,10-Tetraoxa-4,9-diphospha-                                                                10    70  29          2                                     hexacosan-1-aminium, 7-[(hexa-                                                                  10    17              2                                     decyloxy)methyl]-4-hydroxy-9-                                                                   10     0              2                                     methoxy-N,N,N-trimethyl-,                                                                       20    74  26          2                                     hydroxide, inner salt, 4,9-                                                                     20     4              2                                     dioxide, 7(S)-, upper diastereo-                                                                20     0              2                                     mer                                                                           3,5,8,10-Tetraoxa-4,9-diphospha-                                                                10    79  30          2                                     hexacosan-1-aminium, 7-[(hexa-                                                                  10    12              2                                     decyloxy)methyl]-4-hydroxy-9-                                                                   10     0              2                                     methoxy-N,N,N-trimethyl-,                                                                       20    76  28          2                                     hydroxide, inner salt, 4,9-                                                                     20     7              2                                     dioxide, (7R)-, upper                                                                           20     0              2                                     diastereomer      25    93     2.0      2                                     3,5,8,10-Tetraoxa-4,9-diphospha-                                                                10    71  27          2                                     hexacosan-1-aminium, 7-[(hexa-                                                                  10     0              2                                     decyloxy)methyl]-4,9-dihydroxy-                                                                 10     6              2                                     N,N,N-trimethyl-, hydroxide,                                                                    20    72              2                                     4-(inner salt), 4,9-dioxide                                                                     20     4              2                                                       25    91     2.5      2                                     3,5,8,10-Tetraoxa-4,9-diphospha-                                                                10    72  25          2                                     hexacosan-1-aminium, 7-[(hexa-                                                                  10     3              2                                     decyloxy)methyl]-4-hydroxy-9-                                                                   10     0              2                                     methoxy-N,N,N-trimethyl-,                                                                       20    78  39 3.3      2                                     hydroxide, inner salt, 4,9-oxide                                                                20     0              2                                     (7S)-, lower diastereomer                                                                       25    91              2                                     7-(2-Nonadecynyl)-4-hydroxy-                                                                    10    57     4.9      2                                     N,N,N-trimethyl-3,5,9-trioxa-                                                                   20    60              2                                     4-phosphapentacosan-1-aminium,                                                hydroxide, inner salt, 4-oxide                                                7-(2,3-Dioxononadecyl)-4-hydroxy-                                                               10    61     5.4      2                                     N,N,N-trimethyl-3,5,9-trioxa-4-                                                                 20    61              2                                     phosphapentacosan-1-aminium,                                                  hydroxide, inner salt, 4-oxide                                                3,5,8,10-Tetraoxa-4,9-diphospha-                                                                10     5              2                                     pentadecan-1-aminium, 4-hydroxy-                                                                20    11              2                                     9-methoxy-N,N,N-trimethyl-7-                                                  [(pentyloxy)-methyl]-, hydroxide,                                             inner salt, 4,9-dioxide                                                       3,5,8,10-Tetraoxa-4,9-diphospha-                                                                10    20              2                                     pentadecan-1-aminium, 4,9-di-                                                                   20    17              2                                     hydroxy-N,N,N-trimethyl-7-                                                    [(pentyloxy)methyl]-, hydroxide,                                              4-(inner salt), 4,9-dioxide                                                   7-(2-Aminooctadecyl)-4-hydroxy-                                                                 10    95     0.5      2                                     N,N,N-trimethyl-3,5,9-trioxa-4-                                                                 20    95              2                                     phosphapentacosan-1-aminium,                                                  hydroxide, inner salt, 4-oxide                                                3,5,8,10-Tetraoxa-4,9-diphospha-                                                                10    76     5.2      2                                     pentadecan-1-aminium, 7-[(hexa-                                                                 20    83              2                                     decyloxy)methyl]-4-hydroxy-9-                                                 methoxy-N,N,N-trimethyl-,                                                     hydroxide, 4-(inner salt),                                                    4,9-dioxide, less polar isomer                                                3,5,8,10-Tetraoxa-4,9-diphos-                                                                   10    82     2.5      2                                     phapentadecan-1-aminium, 7-                                                                     20    89              2                                     [(hexadecyloxy)methyl]-4-hy-                                                  droxy-9-methoxy-N,N,N-trimethyl-,                                             hydroxide, 4-(inner salt, 4,9-                                                dioxide, more polar isomer                                                    3,5,8,10-Tetraoxa-4,9-diphos-                                                                   10    21              2                                     phapentadecan-1-aminium, 7-                                                                     20    41              2                                     [(hexadecyloxy)methyl]-4,9-                                                   dihydroxy-N,N,N-trimethyl-,                                                   hydroxide, 4-(inner salt),                                                    4,9-dioxide                                                                   3,5,9-Trioxa-4-phosphapenta-                                                                    10    38              2                                     cosan-1-aminium, 7-(3,3-di-                                                                     20    38              2                                     fluoro-2-methylenenonadecyl)-                                                 4-hydroxy-N,N,N-trimethyl-,                                                   hydroxide, inner salt, 4-oxide                                                3,5,9-Trioxa-4-phosphapentacosan-                                                               10    38              2                                     1-aminium, 7-(3,3-difluoro-2-                                                                   20    41              2                                     oxononadecyl)-4-hydroxy-N,N,N-                                                trimethyl-, hydroxide, inner                                                  salt, 4-oxide                                                                 3,5,9-Trioxa-4-phosphapentacosan-                                                               10    45              2                                     1-aminium, 7-(3,3-difluoro-2-hy-                                                                20    49              2                                     droxynonadecyl)-4-hydroxy-N,N,N-                                              trimethyl-, hydroxide, inner                                                  salt, 4-oxide                                                                 7-(2-Aminobutyl)-4,7-dihydroxy-                                                                 10    89     1.3      2                                     N,N,N-trimethyl-3,5,9-trioxa-4-                                                                 20    94              2                                     phosphapentacosan-1-aminium,                                                  hydroxide, inner salt, 4-oxide                                                (2R)-2-[[[(2-Hexadecyl-4-oxo-2-                                                                 10    82     1.4      2                                     azetidinyl)methoxy]hydroxyphos-                                                                 20    79              2                                     phinyl]oxy]-N,N,N-trimethyl-                                                  ethanaminium, hydroxide, inner                                                salt                                                                          7-[(Hexadecylsulfinyl)methyl]-4-                                                                10    23  66 1.4      2                                     hydroxy-N,N,N-trimethyl-3,5,9-                                                                  10    88              2                                     trioxa-4-phosphapentacosan-1-                                                                   10    88              2                                     aminium, hydroxide, inner                                                                       20    93              2                                     salt, 4-oxide                                                                 __________________________________________________________________________

                                      TABLE IV                                    __________________________________________________________________________    HPLA.sub.2                                                                    Compound        Dose μ/ml                                                                        % Inh.                                                                            Avg.                                                                             IC.sub.50 μ/ml                                                                   Avg.                                                                             Subs. con.                              __________________________________________________________________________    3,5,8,10-Tetraoxa-4,9-diphospha-                                                              10    STIM            10                                      hexacosan-1-aminium, 7-[(hexa-                                                                10    STIM            10                                      decyloxy)methyl]-4,9-dihydroxy-                                                               10     0              10                                      N,N,N-trimethyl-hydroxide,                                                                    10     3              10                                      4-(inner salt), 4,9-dioxide                                                                   20    STIM            10                                                      20    STIM            10                                                      20     0              10                                                      20     0              10                                                      50    STIM            10                                                      50    STIM            10                                                      50     0              10                                                      50     0              10                                      3,5,8,10-Tetraoxa-4,9-diphospha-                                                              10    STIM            10                                      hexacosan-1-aminium, 7-[(hexa-                                                                10    STIM            10                                      decyloxy)methyl]-4,9-dihydroxy-                                                               10     0              10                                      N,N,N-trimethyl-hydroxide,                                                                    10     0              10                                      4-(inner salt), 4,9-dioxide                                                                   20    STIM            10                                                      20    STIM            10                                                      20     0              10                                                      20     0              10                                                      50    STIM            10                                                      50    STIM            10                                                      50     0              10                                                      50    STIM            10                                      3,5,8,10-Tetraoxa-4,9-diphospha-                                                              10     1  22          10                                      hexacosan-1-aminium, 7-[(hexa-                                                                10    20              10                                      decyloxy)methyl]-4-hydroxy-9-                                                                 10    36              10                                      methoxy-N,N,N-trimethyl-,                                                                     10    22              10                                      hydroxide, inner salt, 4,9-                                                                   20    18  18          10                                      dioxide, 7(S)-, upper                                                                         20     0              10                                      diastereomer    20    12              10                                                      20    42              10                                                      50    28  23          10                                                      50    34              10                                                      50     8              10                                                      50    23              10                                      3,5,8,10-Tetraoxa-4,9-diphospha-                                                              10    18  17          10                                      hexacosan-1-aminium, 7-[(hexa-                                                                10     0              10                                      decyloxy)methyl]-4-hydroxy-9-                                                                 10    17              10                                      methoxy-N,N,N-trimethyl-,                                                                     10    34              10                                      hydroxide, inner salt, 4,9-                                                                   20    26  21          10                                      dioxide, (7R)-, upper                                                                         20     0              10                                      diastereomer    20    16              10                                                      20    41              10                                                      50    42  30          10                                                      50    23              10                                                      50     9              10                                                      50    44              10                                      3,5,8,10-Tetraoxa-4,9-diphospha-                                                              10     5  18          10                                      hexacosan-1-aminium, 7-[(hexa-                                                                10    14              10                                      decyloxy)methyl]-4,9-dihydroxy-                                                               10     3              10                                      N,N,N-trimethyl-, hydroxide,                                                                  10    48              10                                      4-(inner salt), 4,9-dioxide                                                                   20    16  22          10                                                      20    13              10                                                      20    10              10                                                      20    47              10                                                      50    19  32          10                                                      50    52              10                                                      50     9              10                                                      50    47              10                                      3,5,8,10-Tetraoxa-4,9-diphospha-                                                              10    11  18          10                                      hexacosan-1-aminium, 7-[(hexa-                                                                10    21              10                                      decyloxy)methyl]-4-hydroxy-9-                                                                 10     6              10                                      methoxy-N,N,N-trimethyl-, hy-                                                                 10    32              10                                      droxide, inner salt, 4,9-oxide,                                                               20    14  12          10                                      (7S)-, lower diastereomer                                                                     20     0              10                                                      20     2              10                                                      20    32              10                                                      50    24  25          10                                                      50    15              10                                                      50     8              10                                                      50    52              10                                      7-[(Hexadecylsulfinyl)methyl]-4-                                                              10    27  22          10                                      hydroxy-N,N,N-trimethyl-3,5,9-                                                                10    17              10                                      trioxa-4-phosphapentacosan-1-                                                                 20    33  33          10                                      aminium, hydroxide, inner salt,                                                               20    34              10                                      4-oxide         50    36  42          10                                                      50    47              10                                      __________________________________________________________________________

                                      TABLE V                                     __________________________________________________________________________    MPLA.sub.2                                                                    Compound          Dose μ/ml                                                                        % Inh.                                                                            Avg.                                                                             IC.sub.50 μ/ml                                                                   Avg.                                                                             Subs. con.                            __________________________________________________________________________    N-[3-(Hexadecyloxy)-2-hydroxy-                                                                  25    STIM            2                                     methyl)propyl]octadecanamide                                                                    25    93                                                    4-[(Hexadecyloxy)methyl]-7-                                                                     25    62  79          2                                     hydroxy-N,N,N-triimethyl-1,1-                                                                   25    97              2                                     diphenoxy-2,6,8-trioxa-1,7-di-                                                phosphadecan-10-aminium, hy-                                                  droxide, inner salt, 1,7-dioxide                                              3,5-Dioxa-9-thia-4-phosphapenta-                                                                10    46     7.7      2                                     cosan-1-aminium, 4-hydroxy-N,N,N-                                                               20    83              2                                     trimethyl-, hydroxide, inner                                                                    25    74  80          2                                     salt, 4-oxide     25    86              2                                     7-[(Hexadecylthio)methyl]-4-                                                                    10    59     5.6      2                                     hydroxy-N,N,N-trimethyl-3,5,9-                                                                  20    69              2                                     trioxa-4-phosphapentacosan-1-                                                                   25    79              2                                     aminium, hydroxide, inner salt,                                                                 25    STIM            2                                     4-oxide                                                                       7-[(Hexadecylsulfonyl)methyl]-                                                                  25    80  72          2                                     4-hydroxy-N,N,N-trimethyl-                                                                      25    64              2                                     3,5,9-trioxa-4-phosphapenta-                                                  cosan-1-aminium, hydroxide,                                                   inner salt, 4-oxide                                                           4-Hydroxy-N,N,N-trimethyl-3,5-                                                                  10    24     19.1                                           dioxa-9-thia-4-phosphapentaco-                                                                  20    62                                                    san-1-aminium, hydroxide,                                                     inner salt, 4,9-trioxide                                                      4-Hydroxy-N,N,N-trimethyl-3,5-                                                                  25    73  71          2                                     dioxa-9-thia-4-phosphapentaco-                                                                  25    68              2                                     san-1-aminium, hydroxide, inner                                               salt, 4,9-dioxide                                                             9-Ethoxy-7-[(hexadecyloxy)methyl]-                                                              10    36     16.8     2                                     N,N,N-trimethyl-4-hydroxy-3,5,10-                                                               20    60              2                                     trioxa-4,9-diphosphadodecan-1-                                                                  25    66  78          2                                     aminium, hydroxide, inner salt,                                                                 25    90              2                                     4,9-dihydroxide                                                               4-Hydroxy-N,N,N-trimethyl-7-(2-                                                                 25    STIM            2                                     propanyl)-3,5,9-trioxa-4-phos-                                                                  25    85              2                                     phapentacosan-1-aminium, hydrox-                                              ide, inner salt, 4-oxide                                                      4-Hydroxy-N,N,N-trimethyl-7-                                                                    25    STIM            2                                     (oxiranylmethyl)-35,9-trioxa-4-                                                                 25    85              2                                     phosphapentacosan-1-aminium,                                                  hydroxide, inner salt, 4-oxide                                                4-Hydroxy-7-[2-(hydroxyimino)                                                                   10    19              2                                     ethyl]-N,N,N-trimethyl-3,5,9-                                                                   20    57              2                                     trioxa-4-phosphapentacosan-1-                                                                   25    51  55          2                                     aminium, hydroxide, inner                                                                             60              2                                     salt, 4-oxide                                                                 7-(Boronomethyl)-4-hydroxy-                                                                     25    33  44          2                                     N,N,N-trimethyl-3,5,9-trioxa-                                                                   25    55              2                                     4-phosphapentacosan-1-                                                        aminium, hydroxide, inner salt,                                               4-oxide                                                                       3,5,8,10-Tetraoxa-4,9-diphos-                                                                   25    14  52          2                                     phahexacosan-1-aminium, 7-                                                                      25    89              2                                     [(hexadecyloxy)methyl]-4-hydroxy-                                             9-methoxy-N,N,N-trimethyl-,                                                   hydroxide, inner salt, 4,9-                                                   dioxide                                                                       3,5,8,10-Tetraoxa-4,9-diphos-                                                                   25    STIM            2                                     phahexacosan-1-aminium,                                                                         25    54              2                                     7-[(hexadecyloxy)methyl]-4-                                                   hydroxy-9-methoxy-N,N,N-                                                      trimethyl-, hydroxide, inner                                                  salt, 4,9-dioxide                                                             3,5,8,10-Tetraoxa-4,9-diphospha-                                                                25    28              2                                     hexacosan-1-aminium, 7-[(hexa-                                                                  25     0              2                                     decyloxy)methyl]-4,9-dihydroxy-                                               N,N,N-trimethyl-, hydroxide,                                                  4-(inner salt), 4,9-dioxide                                                   3,5,8,10-Tetraoxa-4,9-diphospha-                                                                25    22              2                                     hexacosan-1-aminium, 7-[(hexa-                                                                  25     8              2                                     decyloxy)methyl]-4,9-dihydroxy-                                               N,N,N-trimethyl-, hydroxide,                                                  4-(inner salt), 4,9-dioxide, (7S)                                             3,5,8,10-Tetraoxa-4,9-diphospha-                                                                25    STIM            2                                     hexacosan-1-aminium, 7-[(hexa-                                                                  25    95              2                                     decyloxy)methyl]-4-hydroxy-9-                                                 methoxy-N,N,N-trimethyl-, hy-                                                 droxide, inner salt, 4,9-                                                     dioxide, 7(S)-, upper                                                         diastereomer                                                                  3,5,8,10-Tetraoxa-4,9-diphospha-                                                                25    STIM            2                                     hexacosan-1-aminium, 7-[(hexa-                                                                  25    93              2                                     decyloxy)methyl]-4-hydroxy-9-                                                 methoxy-N,N,N-trimethyl-,                                                     hydroxide, inner salt, 4,9-                                                   dioxide, (7R)-, upper diastereomer                                            3,5,8,10-Tetraoxa-4,9-diphospha-                                                                25    83              2                                     hexacosan-1-aminium, 7-[(hexa-                                                                  25    91              2                                     decyloxy)methyl]-4-hydroxy-9-                                                 methoxy-N,N,N-trimethyl-,                                                     hydroxide, inner salt, 4,9-                                                   dioxide (7R)-, lower                                                          diastereomer                                                                  3,5,8,10-Tetraoxa-4,9-diphospha-                                                                25    30              2                                     hexacosan-1-aminium, 7-[(hexa-                                                                  25    91              2                                     decyloxy)methyl]-4-hydroxy-9-                                                 methoxy-N,N,N-trimethyl-,                                                     hydroxide, inner salt, 4,9-                                                   oxide, (7S)-, lower diastereomer                                              7-(2-Nonadecynyl)-4-hydroxy-N,N,N-                                                              25    65              2                                     trimethyl-3,5,9-trioxa-4-                                                                       25    STIM            2                                     phosphapentacosan-1-aminium,                                                  hydroxide, inner salt, 4-oxide                                                7-(2,3-Dioxononadecyl)-4-hydroxy-                                                               25    25              2                                     N,N,N-trimethyl-3,5,9-trioxa-4-                                                                 25    STIM            2                                     phosphapentacosan-1-aminium,                                                  hydroxide, inner salt, 4-oxide                                                3,5,8,10-Tetraoxa-4,9-diphospha-                                                                25    STIM            2                                     pentadecan-1-aminium, 4-hydroxy-                                                                25    STIM            2                                     9-methoxy-N,N,N-trimethyl-7-                                                  [(pentyloxy)-methyl]-, hydroxide,                                             inner sale, 4,9-dioxide                                                       3,5,8,10-Tetraoxa-4,9-diphospha-                                                                25    STIM            2                                     pentadecan-1-aminium, 4,9-                                                                      25    STIM            2                                     dihydroxy-N,N,N-trimethyl-7-                                                  [(pentyloxy)-methyl]-, hydroxide,                                             inner sale, 4,9-dioxide                                                       7-(2-Aminooctadecyl)-4-hydroxy-                                                                 10    97  65          2                                     N,N,N-trimethyl-3,5,9-trioxa-4-                                                                 10    34              2                                     phosphapentacosanaminium,                                                                       20    98  85          2                                     hydroxide, inner salt, 4-oxide                                                                  20    72              2                                                       25    92              2                                                       25    STIM            2                                     3,5,8,10-Tetraoxa-4,9-diphospha-                                                                25    33              2                                     pentadecan-1-aminium, 7-[(hexa-                                                                 25    STIM                                                  decyloxy)methyl]-4-hydroxy-9-                                                 methoxy-N,N,N-trimethyl-, hy-                                                 droxide, 4-(inner salt), 4,9-                                                 dioxide, less polar isomer                                                    3,5,8,10-Tetraoxa-4,9-diphos-                                                                   25    61                                                    phapentadecan-1-aminium,                                                                        25    STIM                                                  7-[(hexadecyloxy)methyl]-4-                                                   hydroxy-9-methoxy-N,N,N-                                                      trimethyl-, hydroxide, 4-(inner                                               salt), 4,9-dioxide, more                                                      polar isomer                                                                  3,5,8,10-Tetraoxa-4,9-diphos-                                                                   25    STIM                                                  phapentadecan-1-aminium,                                                                        25    100                                                   7-[(hexadecyloxy)methyl]-4,9-                                                 dihydroxy-N,N,N-trimethyl-,                                                   hydroxide, 4-(inner salt),                                                    4,9-dioxide                                                                   3,5,9-Trioxa-4-phosphapentacosan-                                                               25    80                                                    1-aminium, 7-(3,3-difluoro-2-                                                                   25    STIM                                                  methylenenonadecyl)-4-hydroxy-                                                N,N,N-trimethyl-, hydroxide,                                                  inner salt, 4-oxide                                                           3,5,9-Trioxa-4-phosphapentacosan-                                                               10    88                                                    1-aminium, 7-(3,3-difluoro-2-oxo-                                                               20    89                                                    nonadecyl)-4-hydroxy-N,N,N-tri-                                                                 25    80                                                    methyl-, hydroxide, inner salt,                                                                 25    STIM                                                  4-oxide                                                                       3,5,9-Trioxa-4-phosphapentacosan-                                                               25    82                                                    1-aminium, 7-(3,3-difluoro-2-                                                                   25    STIM                                                  hydroxynonadecyl)-4-hydroxy-                                                  N,N,N-trimethyl-, hydroxide,                                                  inner salt, 4-oxide                                                           7-(2-Aminobutyl)-4,7-dihydroxy-                                                                 10    25     19.2                                           N,N,N-trimethyl-3,5,9-trioxa-4-                                                                 20    61                                                    phosphapentacosan-1-aminium,                                                  hydroxide, inner salt, 4-oxide                                                (2R)-2-[[[(2-Hexadecyl-4-oxo-2-                                                                 10    STIM                                                  azetidinyl)methoxy]hydroxyphos-                                                                 20    STIM                                                  phinyl]oxy]-N,N,N-trimethyl-,                                                 hydroxide, inner salt, 4-oxide                                                3,5,9-Trioxa-4-phosphapentacosan-                                                               10    52     4.9                                            1-aminium, 7-[(bromoacetyl)oxy]-4-                                                              20    85                                                    hydroxy-N,N,N-trimethyl-, hydrox-                                             ide, inner salt, 4-oxide                                                      7-[(Chloroacetyl)oxy]-4-hydroxy-                                                                10    46     9.7                                            N,N,N-trimethyl-3,5,9-trioxa-4-                                                                 20    72                                                    phosphapentacosan-1-aminium,                                                  hydroxide, inner salt, 4-oxide                                                10-Bromo-7-[(hexadecyloxy)methyl]-                                                              10    82     1.5                                            4-hydroxy-N,N,N-trimethyl-9-oxo-                                                                20    82                                                    3,5,8-trioxa-4-phosphahexacosan-1-                                            aminium, hydroxide, inner salt,                                               4-oxide                                                                       7-[(Hexadecylsulfinyl)methyl]-4-                                                                10    71     2.9                                            hydroxy-N,N,N-trimethyl-3,5,9-                                                                  20    84                                                    trioxa-4-phosphapentacosan-1-                                                                   25    98                                                    aminium, hydroxide, inner salt                                                                  25    84                                                    4-oxide                                                                       __________________________________________________________________________

The active compounds of the present invention may be orallyadministered, for example, with an inert diluent, or with an assimilableedible carrier, or they may be enclosed in hard or soft shell capsules,or they may be compressed into tablets, or they may be incorporateddirectly with the food of the diet. For oral therapeutic administration,these active compounds may be incorporated with excipients and used inthe form of tablets, capsules, elixirs, suspensions, syrups and thelike. Such compositions and preparations should contain at least 0.1% ofactive compound. The percentage of the compound in these compositionsmay, of course, be varied and may conveniently be between about 2% toabout 60% of the weight of the unit. The amound of active compound insuch therapeutically useful compositions is such that a suitable dosagewill be obtained. Preferred compositions according to this invention areprepared so that an oral dosage unit contains between about 1 and 250 mgof active compound.

The tablets, capsules and the like may also contain a binder such as gumtragacanth, acacia, corn starch or gelatin; excipients such as dicalciumphosphate; a disintegrating agent such as corn starch, potato starch,alginic acid; a lubricant such as magnesium stearate; and a sweeteningagent such as sucrose, lactose or saccharin. When the dosage unit formis a capsule, it may contain, in addition to materials of the abovetype, a liquid carrier such as a fatty oil.

Various other materials may be present as coatings or to modify thephysical form of the dosage unit. For instance, tablets may be coatedwith shellac, sugar or both. A syrup or elixir may contain, in additionto active ingredient, sucrose as a sweetening agent, methyl andpropylparabens as preservatives, a dye and flavoring such as cherry ororange flavor.

These active compounds may also be administered parenterally. Solutionsor suspensions of these active compounds can be prepared in watersutably mixed with a surfactant such as hydroxypropylcellulose.Dispersions can also be prepared in glycerol, liquid polyethyleneglycols and mixtures thereof in oils. Under ordinary conditions ofstorage and use, these preparations contain a preservative to preventthe growth of microorganisms.

The pharmaceutical forms suitable for injectable use include sterileaqueous solutions or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions. In all cases, the form must be sterile and must be fluid tothe extent that easy syringability exists. It must be stable under theconditions of manufacture and storage and must be preserved against thecontaminating action of microorganisms such as bacterial and fungi. Thecarrier can be a solvent or dispersion medium containing for example,water, ethanol, polyol (e.g. glycerol, propylene glycol and liquidpolyethylene glycol), suitable mixtures thereof, and vegetable oils.

The invention will be further described by the following examples.

EXAMPLE 1 2-[(Hexadecyloxylmethyl]-2-propenoic acid, ethyl ester

To a mixture of 1.1 g of 1-hexadecanol and 400 mg of2-[(acetyloxy)methyl]-2-propenoic acid, ethyl ester in 15 ml ofdichloromethane is added 550 mg of dimethylaminopyridine. This mixtureis stirred overnight, then the solvent is evaporated and the residuesubjected to preparative tlc (25% ethyl acetate/petroleum ether), togive 324 mg of the desired compound.

EXAMPLE 2 3-(Hexadecyloxy)-2-[(hexadecylthio)methyl]propanoic acid,ethyl ester

To a solution of 20 g of 2-[(hexadecyloxy)methyl]-2-propenoic acid,ethyl ester and 10 ml of triethylamine in 60 ml of ethanol is added 53ml of hexadecyl mercaptan. This mixture is stirred overnight, then thesolvents removed and the residue purified by HPLC [hexane:ethyl acetate(9:1)], to give 32.9 g of the desired compound as a white waxy solid.

EXAMPLE 3 3-(Hexadecyloxy)-2-[(hexadecylthio)methyl]-1-propanol

To a slurry of 1.04 g of lithium borohydride in 20 ml of tetrahydrofuranis added 29 g of 3-(hexadecyl-oxy)-2-[(hexadecylthio)methyl]propanoicacid, ethyl ester. This mixture is refluxed overnight, poured into waterand purified by chromatography on silica gel [hexane:ethyl acetate(30:1)], to give 18.4 g of the desired compound.

EXAMPLE 42-Bromoethyl-3-(hexadecyloxy)-2-[(hexadecylthio)methyl]phosphoric acid,propyl ester

To a stirred solution of 15.0 g of3-(hexadecyloxy)-2-[(hexadecylthio)methyl]-1-propanol and 9.6 g of2-bromoethyl phosphorochloridate in 100 ml of carbon tetrachloride isadded 5.7 ml of triethylamine. This mixture is stirred for 3 hours, thenfiltered through diatomaceous earth and the solvents evaporated. Theresidue is dissolved in 25 ml of tetrahydrofuran, 100 ml of 2.5M aqueoussodium acetate added and this mixture is stirred overnight. Diluteaqueous hydrochloric acid and brine are added and this solution wasextracted with ethyl acetate, to give 18.3 g of the desired product, mp.48°-50° C.

EXAMPLE 5 7-[(Hexadecylthio)methyl]-4-hydroxy-N,N,N-trimethyl-3trioxa-4-phosphapentacosan-1-aminium, hydroxide, inner salt, 4-oxide

A mixture of 25.76 g of2-bromoethyl-3-(hexadecyloxy)-2-[(hexadecylthio)methyl]phosphoric acid,propyl ester, 200 ml of acetonitrile, 150 ml of chloroform and 173 ml oftrimethylamine is heated at 60° C. overnight. The solvent is evaporated,150 ml of methanol, 4.0 g of silver carbonate and 3.0 g of ion exchangeresin are added and this mixture is stirred for 2 hours, filtered andthe solvent evaporated. The residue is purified by chromatography, togive 8.5 g of the desired compound as a white solid.

EXAMPLE 6 3-(Hexadecylthio)propanoic acid, ethyl ester

A mixture of 17.7 ml of hexadecyl mercaptan, 3.0 g of ethyl acrylate and2 ml of triethylamine is stirred overnight, the solvent evaporated andthe residue purified by HPLC, to give 19.99 g of the desired compound asa white solid.

EXAMPLE 7 3-(Hexadecylthio)-1-propanol

To a slurry of 147.4 mg of lithium borohydride in ml of tetrahydrofuranis added 2.4 g of 3-(hexadecylthio)propanoic acid, ethyl ester. Thismixture is refluxed overnight, poured into water and extracted withethyl acetate, to give 2.1 g of the desired compound.

EXAMPLE 8 2-Bromoethyl-3-(hexadecylthio)phosphoric acid. propyl ester

To a stirred solution of 2.0 g of 3-(hexadecylthio)-1-propanol and 2.7 gof 2-bromoethyl phosphorochloridate in 25 ml of carbon tetrachloride isadded 1.6 ml of triethylamine. This solution is stirred for 3 hours,filtered through diatomaceous earth and the solvent evaporated. A 25 mlportion of tetrahydrofuran and 25 ml of aqueous 2.5M sodium acetate areadded to the residue. This mixture is stirred overnight, dilute aqueoushydrochloric acid and brine are added and the solution is extracted withethyl acetate, to give 3.0 g of the desired compound, mp. 46°-49° C.

EXAMPLE 97-[(Hexadecylsulfonyl)methyl]-4-hydroxy-N,N,N-trimethyl-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide

A solution of 200 mg of7-[(hexadecylthio)methyl]-4-hydroxy-N,N,N-trimethyl-3,5,9-trioxa-4-phosphapentacosan-1-aminium, hydroxide, inner salt, 4-oxide inchloroform:methanol (1:1) is subjected to ozonolysis at -12° C. at 96volts and a flow rate of 0.5 ml per minute for 25 minutes, to give 204mg of the desired compound.

EXAMPLE 107-[(Hexadecylsulfinyl)methyl]-4-hydroxy-N,N,N-trimethyl-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide

To a solution of 1.0 g of7-[(hexadecylthio)methyl]-4-hydroxy-N,N,N-trimethyl-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide in 5 ml of dichloromethane is added asolution of 323 mg of m-chloroperbenzoic acid in 3 ml ofdichloromethane. This mixture is stirred overnight, then evaporated andthe residue purified by chromatography, to give 1.05 g of the desiredcompound as a white waxy solid.

EXAMPLE 114-Hydroxy-N,N,N-trimethyl-3,5-dioxa-9-thia-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide

A mixture of 3.0 g of 2-bromoethyl-3-(hexadecylthio)phosphoric acid,propyl ester, 36 ml of acetonitrile, 33 ml of chloroform and 20 g oftrimethylamine is heated overnight at 60° C. and then evaporated. To theresidue is added 35 ml of methanol, 1.0 g of silver carbonate and 1.0 gof ion exchange resin. This mixture is stirred for 2 hours, filtered andthe solvent evaporated. The residue is purified by chromatography, togive 1.5 g of the desired compound as a white solid.

EXAMPLE 124-Hydroxy-N,N,N-trimethyl-3,5-dioxa-9-thia-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4.9.9-trioxide

To a solution of 400 mg of4-hydroxy-N,N,N-trimethyl-3,5-dioxa-9-thia-4-phosphapentacosan-l-aminium,hydroxide, inner salt, 4-oxide in 10 1 of dichloromethane containing twodrops of methanol is added 174 mg of m-chloroperbenzoic acid in 3 ml ofdichloromethane. The solution is stirred overnight, then evaporated andthe residue subjected to chromatography, to give 414 mg of the desiredcompound as a white wax.

EXAMPLE 134-Hydroxy-N,N,N-trimethyl-3,5-dioxa-9-thia-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4.9-dioxide

To a solution of 192.4 mg of4-hydroxy-N,N,N-trimethyl-3,5-dioxa-9-thia-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide in 5 ml of chloroform at -78° C. is addeda solution of 68 mg of m-chloroperbenzoic acid in 5 ml of chloroform.This mixture is allowed to warm to room temperature then evaporated. Theresidue is purified by chromatography, to give 173 mg of the desiredcompound as a white wax.

EXAMPLE 142-[(Hexadecyloxy)methyl]-3-(phenylmethoxy)-4-methylbenzenesulfonate-1-propanol

To a solution of 23 g of3-[(hexadecyloxy)methyl]-4-(phenylmethoxy)-butanol and 25 ml oftriethylamine in 50 ml of dichloromethane at 0° C., is added 11.633 g ofp-toluenesulfonyl chloride in 20 ml of dichloromethane. This solution isstirred overnight and then poured into water. The organic layer isseparated, then evaporated and purified by chromatography, to give 31 gof the desired compound as a dark yellow oil.

EXAMPLE 15 [[(3-Hexadecyloxy)-2-(iodomethyl)propoxy1methyl]benzene

A mixture of 5.3g of3-[(hexadecyloxy)methyl]-4-(phenylmethoxy)-4-toluenesulfonyloxybutane,4.2 g of sodium iodide and 50 ml of acetone is stirred at reflux for 2days, then poured into water and extracted with dichloromethane. Theextract is washed with saturated aqueous sodium bisulfite, dried andevaporated. The residue is purified by chromatography, to give 3.5 g ofthe desired compound as a light yellow oil.

EXAMPLE 16 2-[(Hexadecyloxy)methyl]-3-(phenylmethoxy)propyl]phosphonicacid, diethyl ester

To 20 mg of washed sodium in 3 ml of toluene under nitrogen is added0.118 ml of diethyl phosphite. This mixture is stirred for 2 hours, thenheated to 80° C. for 30 minutes, giving a clear solution. To thissolution is added 200 mg of[[(3-hexadecyloxy)-2-(iodomethyl)propoxy]methyl]benzene in 1 ml oftoluene at 80° C. This mixture is heated at 80° C. overnight, then thesolvent is evaporated. Water and ether are added to the residue. Theether layer is separated, dried and evaporated. This residue is purifiedby chromatography, giving 117 mg of the desired compound as a clear oil.

EXAMPLE 17 [3-(Hexadecyloxy)-2-(hydroxymethyl)propyl]phosphonic acid,diethyl ester

A 7.7 g portion of2-[(hexadecyloxy)methyl]-3-(phenylmethoxy)propyl]phosphonic acid,diethyl ester is hydrogenated over 500 mg of 10% palladium-on-carbon in20 ml of methanol and 20 ml of acetic acid, to give 6.5 g of the desiredcompound.

EXAMPLE 18 2-Bromomethyl-3-(diethoxyphosphinyl)-2-((hexadecyloxy)methyl]phosphoric acid, propyl ester

To 5.0 g of [3-(hexadecyloxy)-2-(hydroxymethyl) propyl]phosphonic acid,diethyl ester in 50 ml of carbon tetrachloride is added 5.0 g of2-bromomethyl phosphorodichloridate, followed by 3 ml of triethylamine.This mixture is stirred for 2.5 hours, then filtered throughdiatomaceous earth and the solvent evaporated. The residue is mixed with50 ml of tetrahydrofuran and 50 ml of aqueous 0.5N sodium acetate andstirred overnight. Dilute hydrochloric acid, brine and ethyl acetate areadded. The organic layer is separated, dried and evaporated, to give 6.8g of the desired compound as a yellow oil.

EXAMPLE 19 9-Ethoxy-7-[(hexadecyloxy)methyl]-N,N,N-trimethyl-4-hydroxy-3,5,10-trioxa-4,9-diphosphadodecan-1-aminium. hydroxide, innersalt, 4,9-dihydroxide

A mixture of 6.8 g of2-bromoethyl-3-(diethoxyphosphinyl)-2-[(hexadecyloxy)methyl]phosphoricacid, propyl ester, 30 ml of acetonitrile, 30 ml of chloroform and 5.0 gof trimethylamine is mixed in a glass bomb and heated overnight at 60°C. The solvent is evaporated, 100 ml of methanol and 1.0 of silvercarbonate added and this mixture stirred at room temperature for 2hours. The mixture is filtered through diatomaceous earth and thenevaporated, to give a white solid which is purified by chromatography toyield 4.4 of the desired compound as a white wax.

EXAMPLE 20 2-[(Hexadecyloxy)methyl]-2-propen-1-ol

To 18.5 g of washed sodium hydride in 500 ml of dry dimethylformamide ina water bath is added dropwise, 67.4 g of 2-butene-1,2-diol. A solutionof 235 g of hexadecylbromide in 100 ml of dimethylformamide is addeddropwise. This mixture is stirred at room temperature for 2 days thenpoured into water and extracted with ether. The ether extract isseparated, dried, evaporated and purified by chromatography, to give 30g of the desired compound.

EXAMPLE 21(1,1-Dimethylethyl)[[2-[(hexadecyloxy)methyl]-2-propenyl]oxy]dimethylsilane

To a solution of 5.0 g of 2-[(hexadecyloxy)-methyl]-2-propen-1-ol in 50ml of dry N,N-dimethylformamide is added 6.05 g oftert-butyldimethylsilyl chloride followed by 8 g of imidazole. Thismixture is stirred overnight, then diluted with ether, washed withsaturated aqueous sodium bicarbonate, water and brine, dried andevaporated, to give 6.49 g of the desired compound.

EXAMPLE 225-[[[(1.1-Dimethylethyl)dimethylsilyl]oxy]methyl]-3-ethyl-5-[(hexadecyloxy)methyl]-4.5-dihydroisoxazole

To a solution of 2.0 g of(1,1-dimethylethyl)[[2-[(hexadecyloxy)methyl]-2-propenyl]oxy]dimethylsilane,623 mg of propyl nitrite and 3 ml of triethylamine in a water bath isslowly added a solution of 1.07 g of phosphorous oxychloride in 2 ml ofchloroform. This solution is warmed to room temperature, stirredovernight, then poured into water, dichloromethane and brine. Theorganic layer is separated, dried and evaporated. The residue issubjected to flash chromatography, to give 1.0 g of the desiredcompound.

EXAMPLE 23(S)-3-Ethyl-4,5-dihydro-5-[(hexadecyloxy)methyl]-5-isoxazolemethanol

To a solution of 500 mg of5-[[[(1,1-dimethylethyl)dimethylsilyl]oxy]methyl]-3-ethyl-5-[(hexadecyloxy)methyl]-4,5-dihydroisoxazolein 10 ml of tetrahydrofuran is added 2.13 ml of 1M tetrabutylammoniumfluoride in tetrahydrofuran. After 2 hours the reaction is poured intowater, ether and brine. The organic layer is separated, dried andevaporated, to give 300 mg of the desired compound.

EXAMPLE 24(5S)-2-[[[[3-Ethyl-5-[(hexadecyloxy)methyl]-4,5-dihydro-5-isoxazolyl]methoxy]hydroxyphosphinyl]oxy]-N,N,N-trimethylethanaminium,hydroxide, inner salt

To a solution of 1.1 g of(S)-3-ethyl-4,5-dihydro-5-[(hexadecyloxy)methyl]-5-isoxazolemethanol in15 ml of dry carbon tetrachloride under argon is added 1.02 g of2-bromoethyl phosphorochloridate and 5 ml of triethylamine. This mixtureis stirred for 2 hours, filtered through diatomaceous earth and thesolvent evaporated. To the residue is added 5 ml of tetrahydrofuran and5 ml of 0.5N aqueous sodium acetate. This mixture is stirred overnight.Dilute hydrochloric acid and ethyl acetate are added, the organic layerseparated, washed with brine, dried and evaporated. The residue isplaced in a bomb with 21 ml of acetonitrile, 21 ml of chloroform and 20g of trimethylamine and heated at 65° C. for 5 hours. The solvents areevaporated. Methanol and silver carbonate are added, the mixture stirredfor 2 hours, filtered and evaporated, to give, after chromatography, 900mg of the desired compound.

EXAMPLE 257-(2-Aminobutyl)-4,7-dihydroxy-N,N,N-trimethyl-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide

A mixture of 500 mg of(5S)-2-[[[[3-ethyl-5-[(hexadecyloxy)methyl]-4,5-dihydro-5-isoxazolyl]methoxy]hydroxyphosphinyl]oxy]-N,N,N-trimethylethanaminium,hydroxide, inner salt, 50 mg of 10% palladium-on-carbon and 15 ml ofmethanol is hydrogenated for 20 hours, then filtered and evaporated, togive 500 mg of the desired compound as a clear oil.

EXAMPLE 26 2-[(Phenylmethoxy)methyl]-1-octadecene

To 1.4 g of washed sodium hydride in 100 ml of dry N,N-dimethylformamideis added a solution of 10 g of 1-octadecenol in 100 ml ofdimethylformamide followed by 5.8 g of benzyl bromide. This mixture isstirred for 15 hours, then water added and the mixture extracted withether. The ether extract is dried, evaporated and purified bychromatography, to give 9.0 g of the desired compound.

EXAMPLE 27 (R)-4-Hexadecyl-4-[(phenylmethoxy)methyl]-2-azetidinone

To a solution of 3.0 g of 2-[(phenylmethoxy)methyl]-1-octadecene in 10ml of benzene under argon is added 1.05 ml of chlorosulfonyl isocyanate.The mixture is stirred overnight and then poured into a solution of 22 gof sodium bisulfite, 100 ml of water, 5 ml of 10% sodium hydroxide and100 ml of ether. This mixture is stirred for 2 hours, and the organiclayer separated, dried and evaporated. The residue is purified bychromatography, to give 1.1 g of the desired compound as a colorlessoil.

EXAMPLE 28 (R)-4-Hexadecyl-4-(hydroxymethyl)-2-azetidinone

A mixture of 400 mg of(R)-4-hexadecyl-4-[(phenylmethoxy)methyl]-2-azetidinone, 50 mg of 10%palladium-on-carbon and 20 ml of methanol:acetic acid (1:1) ishydrogenated for 16 hours, then filtered and evaporated. The residue isrecrystallized from petroleum ether/ethyl acetate, to give 300 mg of thedesired compound as a white solid, mp. 82° C.

EXAMPLE 29(2R)-2-[[[(2-Hexadecyl-4-oxo-2-azetidinyl)methoxy]hydroxyphosphinyl]oxy]-N,N,N-trimethylethanaminium,hydroxide, inner salt

To a mixture of 265 mg of(R)-4-hexadecyl-4-(hydroxymethyl)-2-azetidinone and 86 mg oftriethylamine in 4 ml of dry benzene at 0° C. under argon is added 121mg of 2-chloro-2-oxo-1,3,2-dioxaphospholine. The mixture is warmed toroom temperature, stirred overnight, filtered through diatomaceous earthand evaporated. The residue is dissolved in 15 ml of dry acetonitrile, 1ml of trimethylamine is added and this mixture placed in a bomb andheated overnight at 65° C. The product is purified by chromatography, togive 90 mg of the desired compound as a white solid.

EXAMPLE 30 2-Nonadecyn-1-ol

A mixture of 56.1 g of propargyl alcohol in 100 ml of tetrahydrofuranand 15.6 g of lithium in 1 liter of ammonia containing a trace of ferricchloride is stirred for 1 hour. A 76.4 ml portion of hexadecyl bromideis added and the mixture refluxed for 8 hours then evaporated. Theresidue is dissolved in ether and washed with 10% hydrochloric acid,dried and purified by flash chromatography, to give 47.4 g of thedesired compound as white platelets, mp. 60° C.

EXAMPLE 31 1-Bromo-2-nonadecyne

A mixture of 18.8 g of 2-nonadecyn-1-ol, 110 ml of ether, 1.14 ml ofpyridine and 5.7 ml of phosphorous tribromide in 25 ml of ether isheated at reflux for 2 hours, then stirred at room temperature for 2hours, poured into water and washed with hydrochloric acid. The solutionis evaporated and filtered through silicon dioxide with hexane, to give16.9 g of the desired product as a white solid, mp. 34°-35° C.

EXAMPLE 32 2-Nonadecyvylpropanedioic acid, diethyl ester

A mixture of 16.5 g of 1-bromo-2-nonadecyne in 50 ml of ethanol is addedto 3.0 g of 60% sodium hydride and 11.4 ml of diethyl malonate in 100 mlof ethanol. This mixture is stirred one hour and then evaporated. Theresidue is washed with ether, twice with dilute hydrochloric acid andbrine. The residue is then purified by flash chromatography, (5% ethylacetate:petroleum ether), to give 12.1 g of the desired compound aswhite needles, mp. 33°-34° C.

EXAMPLE 33 2-(2-Nonadecynyl)-1,3-propanediol

A mixture of 1.8 g of lithium aluminum hydride in 150 ml of ether isprepared at 0° C. To this is added a solution of 11.7 g of2-nonadecynylpropanedioic acid, diethyl ester in ether. This mixture isrefluxed for 4 hours then the reaction is quenched with aqueous sodiumsulfate. After stirring for 2 days, the reaction is diluted with ethylacetate and filtered. The residue is treated with methanol, IN sodiumhydroxide and tetrahydrofuran and then evaporated. The residue isdissolved in ether and washed with brine and purified by flashchromatography, (4:6 ethyl acetate:petroleum ether), to give 5.3 g ofthe desired product as white plates, mp. 55° C.

EXAMPLE 34 2-[(Hexadecyloxy)methyl]-4-heneicosyn-1-ol

A mixture of 5.0 g of 2-(2-nonadecynyl)-1,3-propanediol, 75 ml ofN,N-dimethylformamide, 710 mg of 60% sodium hydride and 5.4 ml ofhexadecyl bromide is heated at 70° C. for 2 hours and then evaporated.The residue is purified by flash chromatography with an ethylacetate:petroleum ether gradient to give 5.8 g of the desired compoundas white crystals, mp. 46°-47° C.

EXAMPLE 357-(2-Nonadecynyl)-4-hydroxy-N,N,N-trimethyl-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide

A mixture of 4.0 g of 2-[(hexadecyloxy)methyl]-4-heneicosyn-1-ol, 50 mlof carbon tetrachloride, 1.6 ml of triethylamine and 1.38 ml of2-bromoethyl phosphorochloridate is stirred for 4 hours at roomtemperature and the solvent evaporated. The residue is diluted with 100ml of tetrahydrofuran and 100 ml of 0.5M sodium acetate and stirredovernight, then evaporated. This residue is acidified and extractedthree times with ether. The extracts are combined, evaporated and theresidue added to a mixture of 50 ml of chloroform, 50 ml of acetonitrileand 25 g of trimethylamine. The mixture is heated at 60° C. for 4.5hours, then evaporated and purified by flash chromatography (3:7 methylalcohol: chloroform with 5% water), to give 4.2 g of the desiredcompound as an amorphous solid.

EXAMPLE 36 7-(2,3-Dioxononadecyl)-4-hydroxy-N,N,N-trimethyl-3,5,9-trioxa-4-phosphapentacosan-1-aminium, hydroxide, inner salt, 4-oxide

A mixture of 500 mg of 7-(2-nonadecynyl)-4-hydroxy-N,N,N-trimethyl-3,5,9-trioxa-4-phosphapentacos-an-aminium,hydroxide, inner salt, 4-oxide, 5 ml of dichloromethane and 0.25 ml ofacetic acid is heated at reflux. A 100 mg portion of tricaprylylmethylammonium chloride is added followed by 500 mg of ground potassiumpermangamate. The reaction is heated at reflux for 1 hour. The crudereaction mixture is subjected to flash chromatography (3:7 methylalcohol:chloroform with 5% water) to give 135 mg of the desired compoundas a yellow amorphous solid.

EXAMPLE 37 2-Methylene-1-octadecanol

This compound is prepared by the process of Marshall, Journal of OrganicChemistry, 32, 113 (1967).

EXAMPLE 38 2-(Bromomethyl)-1-octadecene

A mixture of 20 g of 2-methylene-1-octadecanol, 150 ml of ether and 12.6ml of pyridine is cooled to -15° C. in an ice-methanol bath. A solutionof 7.3 ml of phosphorous tribromide in 50 ml of ether is added slowlyover 1 hour, and the mixture is allowed to warm to room temperatureovernight. The mixture is recooled to 0° C., 1.9 ml of water added andthen allowed to warm to room temperature over 6 hours. The mixture iswashed, filtered and evaporated, to give 19.2 g of the desired compoundas a clear oil.

EXAMPLE 39 2-(Methyleneoctadecyl)propanedioic acid, diethyl ester

A mixture of 3.7 g of 50% sodium hydride, 100 ml of tetrahydrofuran and10.1 ml of diethylmalonate is cooled with a water bath. A solution of19.2 g of 2-(bromomethyl)-1-octadecene in 50 ml of tetrahydrofuran isadded dropwise and this mixture stirred overnight at room temperature. A2.6 g portion of 50% sodium hydride is added and the mixture refluxedovernight, then diluted with ether and washed with dilute hydrochloricacid, water and brine. Flash chromatography (5% ethyl acetate:hexane)gives 8.52 g of the desired compound as a clear oil.

EXAMPLE 40 2-(2-Methyleneoctadecyl)-1,3-propanediol

A solution of 1.03 g of lithium aluminum hydride in 50 ml of ether iscooled in a water bath. A mixture of 6.78 g of2-(methyleneoctadecyl)propanedioic acid, diethyl ester in 50 ml of etheris added. This mixture is stirred at reflux for 2 hours, then overnightat room temperature and finally at reflux for 4 hours. The mixture isstirred with sodium sulfate decahydrate, then filtered andrecrystallized from ethyl acetate/hexane, to give 4.87 g of the desiredcompound as white crystals, mp. 64°-65° C.

EXAMPLE 41 2-[(Hexadecyloxy)methyl]-4-methylene-1-eicosanol

A solution of 80 mg of 60% sodium hydride in 20 ml ofN,N-dimethylformamide is stirred at room temperature. A 680 mg portionof 2-(2-methyleneoctadecyl)-1,3-propanediol and 0.61 ml ofhexadecylbromide are added and the mixture warmed very slowly to 60°-70°C. The mixture is evaporated, dissolved in ether, and is washed withwater and brine then flash chromatographed (5% ethyl acetate:petroleumether), to give 550 mg of the desired compound as colorless crystals,mp. 37°-38° C.

EXAMPLE 424-Hydroxy-N,N,N-trimethyl-7-(2-methyleneoctadecyl)-3,5,9-trioxa-4-phosphaoentacosan-1-aminium,hydroxide, inner salt, 4-oxide

A mixture of 10.2 g of 2-[(hexadecyloxy)methyl]-4-methylene-1-eicosanol,100 ml of carbon tetrachloride, 4.0 ml of triethylamine and 6.41 g of2-bromoethyl phosphorodichloridate is stirred for 3 hours, then filteredand evaporated. To the residue is added 200 ml of tetrahydrofuran and200 ml of 0.5M aqueous sodium acetate. This mixture is stirred for 2hours, the tetrahydrofuran evaporated and the residue acidified withhydrochloric acid and extracted with ether. The extract is dried andevaporated. The residue is combined with 150 ml of acetonitrile, 150 mlof chloroform and 50 g of trimethylamine and refluxed for 4 hours. Thesolution is evaporated and the residue stirred with 200 ml each ofmethanol and tetrahydrofuran and 2.5 g of silver carbonate for 90minutes, then filtered and evaporated. The residue is purified by flashchromatography (30% methyl alcohol:chloroform up to 5% water:30% methylalcohol:chloroform), to give 10.1 g of the desired compound as anamorphous white solid.

EXAMPLE 434-Hydroxy-N,N,N-trimethyl-7-(2-oxooctadecyl)-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide inner salt, 4 oxide

A mixture of 5.0 g of4-hydroxy-N,N,N-trimethyl-7-(2-methyleneoctadecyl)-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide, 200 ml of chloroform and 22 ml ofethanol is subjected to ozonolysis at -40° C. until the solution turnsblue. The reaction is flushed with oxygen, 1.6 ml of trimethyl phosphiteadded at 0° C. and then evaporated. The residue is purified by flashchromatography (30% methyl alcohol:chloroform up to 5% water:30% methylalcohol:chloroform), to give 4.0 g of the desired compound as anamorphous white solid.

EXAMPLE 447-(2-Aminooctadecyl)-4-hydroxy-N,N,N-trimethyl-3,5,9-trioxa-4-phosphapentacosanaminium,hydroxide, inner salt, 4-oxide

A mixture of 200 mg of4-hydroxy-N,N,N-trimethyl-7-(2-methyleneoctadecyl)-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide, 2 ml of methanol, 1 ml of chloroform,300 mg of ammonium acetate and 15 mg of sodium cyanoborohydride isstirred for 36 hours and then evaporated and chromatographed (30% methylalcohol:chloroform up to 5% water:30% methyl alcohol:chloroform, up to5% ammonium hydroxide:30% methyl alcohol:chloroform), to give 160 mg ofthe desired compound as a white amorphous solid.

EXAMPLE 454-Hydroxy-7-(2-hydroxy-2-methyloctadecyl)-N,N,N-trimethyl-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide

A mixture of 410 mg of4-hydroxy-N,N,N-trimethyl-7-(2-oxooctadecyl)-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide, 10 ml of tetrahydrofuran, 5 ml oftoluene and 1.5 ml of hexamethyl phosphoramide is stirred at 0° C. A 1ml portion of 3M methyl magnesium bromide is added in portions, and themixture is stirred at room temperature overnight. The mixture isrecooled to 0° C., 1 ml of methyl magnesium bromide, 0.6 ml of aceticacid in tetrahydrofuran and an excess of sodium bicarbonate is addedsequentially and the mixture is evaporated. The residue is purified bychromatography (3:7 methyl alcohol:chloroform up to 5% water in 3:7methyl alcohol:chloroform), to give 200 mg of the desired compound as anamorphous white solid.

EXAMPLE 467-[(2-Hexadecyloxiranyl)methyl]-4-hydroxy-N,N,N-trimethyl-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide

A mixture of 290 mg of 3-chloroperoxybenzoic acid, 1.3 g of sodiumbicarbonate and 15 ml of chloroform is stirred rapidly at 0° C. A 730 mgportion of4-hydroxy-N,N,N-trimethyl-7-(2-methyleneoctadecyl)-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide is added and this mixture is stirred atroom temperature for 5 hours, then at 0° C. overnight. A 75 ml portionof methyl sulfide is added and the mixture flash chromatographed (3:7methyl alcohol:chloroform up to 5% water in 3:7 methylalcohol:chloroform), to give 700 mg of the desired compound as anoff-white amorphous solid.

EXAMPLE 474-Hydroxy-7-(2-hydroxyoctadecyl)-N,N,N-trimethyl-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide and isomer

A mixture of 732 mg of4-hydroxy-N,N,N-trimethyl-7-(2-oxooctadecyl)-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide and 10 ml of chloroform is stirred at 0°C. A 25 ml portion of diborane in tetrahydrofuran is added, and themixture stirred at room temperature then evaporated. The residue isflash chromatographed (3:7 methyl alcohol:chloroform up to 5% water in3:7 methyl alcohol:chloroform), to give 210 mg and 150 mg of therespective isomeric forms as white amorphous solids.

EXAMPLE 48 [[3-Azido-2-[(hexadecyloxy)methyl]propoxy]methyl]benzene

A mixture of 16.3 g of2-[(hexadecyloxy)methyl]-3-(phenylmethoxy)-4-methylbenzenesulfonate-1-propanol,9.22 g of sodium azide and 130 ml of N,N-dimethylformamide is stirred at80° C. for 3.5 hours and then at room temperature overnight. The mixtureis poured into water and extracted with ether. The ether layer is driedand the solvent removed. The residue is purified by chromatography, togive 8.9 g of the desired compound as an oil.

EXAMPLE 49 2-[(Hexadecyloxy)methyl]-3-(phenylmethoxy)-1-propanamine

To a solution of 0.87 g of lithium aluminum hydride in 80 ml of etherunder argon is added, with stirring, over 30 minutes, a solution of 8.5g of [[3- azido-2-[(hexadecyloxy)methyl]propoxy]methyl]benzene in 80 mlof ether. This mixture is refluxed for 3 hours, then cooled to 0° C. and8 ml of saturated aqueous sodium sulfate solution is added. The mixtureis filtered and the filtrate evaporated, to give 8.0 g of the desiredcompound as an oil.

EXAMPLE 50 N-[2-[(Hexadecyloxy)methyl]-3-(phenylmethoxy)propyl]octadecanamide

A mixture of 8.0 g of2-[(hexadecyloxy)methyl]-3-(phenylmethoxy)-1-propanamine, 12.6 g ofstearic anhydride, 2.56 g of dimethylaminopyridine and 30 ml ofchloroform is stirred overnight, and the solvent evaporated. Ether isadded to the residue and this solution washed with dilute hydrochloricacid and saturated aqueous sodium bicarbonate. The solvent is removedand the residue purified by chromatography and then recrystallized frommethanol, to give 11.0 g of the desired compound as a white solid, mp.56°-57° C.

EXAMPLE 51 N-[3-(Hexadecyloxy)-2-hydroxymethyl)propyl]octadecanamide

A mixture of 10.5 g ofN-[2-[(hexadecyloxy)methyl]-3-(phenylmethoxy)propyl]octadecanamide, 1.5g of 5% palladium-on-carbon, 250 ml of cyclohexane and 27 ml of glacialacetic acid is shaken in a Parr apparatus for 24 hours and thenfiltered. The filtrate is evaporated and the residue recrystallized frommethanol, to give 7.9 g of the desired product as a white solid, mp.74°-75° C.

EXAMPLE 527-[(Hexadecyloxy)methyl]-4-hydroxy-N,N,N-trimethyl-10-oxo-3,5-dioxa-9-aza-4-phosphaheptacosan-1-aminium,hydroxide, inner salt, 4-oxide

A mixture of 5.0 g ofN-[3-(hexadecyloxy)-2-hydroxymethyl)propyl]octadecanamide, 1.31 g of2-chloro-1,3,2-dioxaphospholane-2-oxide, 1.02 g of triethylamine and 200ml of ether is stirred overnight, then diluted with more ether and ethylacetate, washed with brine and dried. The solvent is removed and theresidue heated at 65° C. with 50 ml of chloroform, 100 ml ofacetonitrile and 60 ml of 33% trimethylamine in acetonitrile in apressure bottle overnight. An additional 40 ml of trimethylamine isadded, the mixture is heated at 70° C. for 30 minutes and then thesolvent is removed. The residue is purified by chromatography, to give2.3 g of the desired compound.

EXAMPLE 53 2-[(Hexadecyloxy)methyl]-4-penten-1-ol

To a suspension of 19.1 g of 50% sodium hydride under argon in 300 ml ofN,N-dimethylformamide is added dropwise over a period of 1 hour, withstirring a solution of 37 g of 4-penten-1-ol, 2-methanol (prepared asdescribed by B. K. Wasson, et. al., J. Chem. Soc., 39, 923 (1961)) in200 ml of tetrahydrofuran. After stirring another 30 minutes, 126.34 gof hexadecyloxytosylate is added followed by 500 ml of tetrahydrofuran.This mixture is stirred for 2.5 hours, then heated to 80° C. for 20minutes and then at room temperature overnight. Water is added and themixture extracted with ether. The ether extract is dried and thenevaporated. The residue is distilled using a Kugelrohr. The fractiondistilling at 200°-210° C. 1 mm is collected, purified by chromatographyand then redistilled, to give 11.35 g of the desired compound.

EXAMPLE 544-Hydroxy-N,N,N-trimethyl-7-(2-propanyl)-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide

A mixture of 10.3 g of 2-[(hexadecyloxy)methyl]-4-penten-1-ol, 9.14 g of2-bromoethyl phosphorochloridate, 3.83 g of triethylamine, and 225 ml ofcarbon tetrachloride is stirred for 10 minutes and then refrigeratedovernight. The mixture is stirred with 250 ml of tetrahydrofuran and 250ml of 0.5M aqueous sodium acetate for 2 hours and most of thetetrahydrofuran is removed. The residue is acidified with hydrochloricacid and extracted with ether. The ether extract is dried andevaporated. The residue is refluxed in a mixture of 150 ml ofacetonitrile, 120 ml of chloroform and 50 g of trimethylamine for 4hours and then evaporated. The residue is dissolved in 200 ml ofmethanol and stirred with 20 g of ion exchange resin and 2 g of silvercarbonate for 1 hour then filtered and evaporated. The residue ispurified by chromatography to give 10.2 g of the desired compound as acolorless solid.

EXAMPLE 554-Hydroxy-N,N,N-trimethyl-7-(2-oxoethyl)-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide

A solution of 2.0 g of4-hydroxy-N,N,N-trimethyl-7-(2-propenyl)-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide in 35 ml of methanol and 25 ml ofdichloromethane is stirred at -78° C. as ozone was bubbled through for30 minutes. The mixture is allowed to warm to -40° C., then recooled to-78° C. and 0.98 g of trimethylphosphite is added. The mixture isallowed to warm to room temperature and the solvent removed. The residueis purified by chromatography, to give 1.0 g of the desired compound asa white powder.

EXAMPLE 564-Hydroxy-7-[2-(hydroxyimino)ethyl]-N,N,N-trimethyl-3,5,9-trioxa-4-phosphaoentacosan-1-aminium,hydroxide, inner salt, 4-oxide

A mixture of 0.45 g of4-hydroxy-N,N,N-trimethyl-7-(2-oxoethyl)-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide, 0.12 g of hydroxylamine hydrochloride,0.055 g of sodium carbonate, 6 ml of ethanol and 3 ml of water isstirred for 40 minutes and then refrigerated overnight. Another 0.12 gof hydroxylamine hydrochloride and 0.05 g of sodium carbonate are addedand the mixture stirred for 4 hours. The mixture is diluted withethanol, passed through a column of ion exchange resin and the solventremoved. The residue is dissolved in chloroform, filtered throughdiatomaceous earth and evaporated. The residue is triturated with etherand the solid collected, to give 0.45 g of the desired compound as ahydroscopic white powder.

EXAMPLE 574-Hydroxy-N,N,N-trimethyl-7-(oxiranylmethyl)-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide

A mixture of 3.0 g of4-hydroxy-N,N,N-trimethyl-7-(2-propenyl)-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide, 2.25 g of m-chloroperbenzoic acid and 30ml of dichloromethane is stirred for 24 hours and then refrigerated for18 hours. The solvent is removed and the residue purified bychromatography, then triturated with ether, to give 1.8 g of the desiredcompound as a white powder.

EXAMPLE 582-[[Hydroxy[(2-methylenenonadecyl)oxy]phosphinyl]oxy]-N,N,N-trimethylethaniminium,hydroxide, inner salt

To a solution of 3.8 g of 2-[(hexadecyloxy)methyl]-2-propen-1-ol in 100ml of carbon tetrachloride is added 3.91 g of 2-bromoethylphosphorochloridate and 1.64 g of triethylamine. The mixture is stirredovernight in a refrigerator, then filtered and the solvent removed. A100 ml portion of 0.5M aqueous sodium acetate and 100 ml oftetrahydrofuran are added and this mixture stirred for 1 hour. Thetetrahydrofuran is removed, the mixture acidified with sulfuric acid andextracted with ether. The ether extract is washed with brine, dried andevaporated. The residue is stirred at reflux in 60 ml of acetonitrile,50 ml of chloroform and 20 g of trimethylamine for 4 hours, thenevaporated. The residue is stirred in 100 ml of methanol containing 10 gof ion exchange resin and 1.0 g of silver carbonate, then filtered andthe solvent removed. This residue is purified by chromatography, to give2.4 g of the desired compound as a white solid.

EXAMPLE 597-(Boronomethyl)-4-hydroxy-N,N,N-trimethyl-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide

To a stirred solution of 46 ml of 0.98 molar borane in tetrahydrofuranat 0° C. under argon is added over 20 minutes, a solution of 2.4 q of2-[[hydroxy[(2methylenenonadecyl)oxy]phosphinyl]oxy]-N,N,N-trimethylethaniminium,hydroxide, inner salt in 75 ml of dry dichloromethane. This mixture isstirred for 1.5 hours at 0° C., then the excess tetrahydrofuran andborane are removed. A 50 ml portion of tetrahydrofuran:water (10:1) isadded and the solvent removed. Methanol is added and evaporated. Theresidual oil is chromatographed and the product triturated with ether,to give 0.72 g of the desired product as a white powder.

EXAMPLE 60 2-[(Hexadecyloxy]methyl]-3-(phenylmethoxy)propyl phosphoricacid, diphenyl ester

To a solution of 1.0 g of3-[(hexadecyloxy)methyl]-4-(phenylmethoxy)-butanol in a mixture of 10 mlof dry acetonitrile and 10 ml of dry dichlormethane is added 831 mg of5-chloro-1-methyl imidazole followed by 830 mg of diphenylchlorophosphate. This mixture is stirred overnight under argon. An 830mg portion of diphenyl chlorophosphate is added and stirring continued.The solvent is removed and the residue purified by chromatography, togive 1.27 g of the desired compound as a colorless oil.

EXAMPLE 61 3-(Hexadecyloxy)-2-(hydroxymethyl)propyl phosphoric acid,diphenyl ester

A solution of 1.27 g of 2-[(hexadecyloxy)methyl]-3-(phenylmethoxy)propylphosphoric acid, diphenyl ester in 50 ml of cyclohexane and 5 ml ofglacial acetic acid is hydrogenated over 500 mg of 5%palladium-on-carbon in a Parr apparatus at an initial pressure of 25 psifor 18 hours. The mixture is filtered and washed with cyclohexane. Thecombined filtrate and wash is evaporated to give an oil. This oil isevaporated twice from toluene, to give 685 mg of the desired compound asan oil.

EXAMPLE 624-[(Hexadecyloxy)methyl]-7-hydroxy-N,N,N-trimethyl-1,1-diphenoxy-2.6.8trioxa-1.7-diophosphadecan-10-aminium, hydroxide, inner salt,1,7-dioxide

To a solution of 0.65 g of 3-(hexadecyloxy)-2-(hydroxymethyl)propylphosphoric acid, diphenyl ester in 15 ml of carbon tetrachloride,stirred in an ice bath under argon is added 0.48 ml of triethylaminefollowed by the dropwise addition of 0.42 g of phosphoryl dichloride in2 ml of carbon tetrachloride. This mixture is stirred overnight at roomtemperature, then 10 ml of toluene is added and the mixture is filteredand washed with toluene. The filtrate and wash are combined and taken todryness. The residue is stirred in 50 ml of tetrahydrofuran and 50 ml of0.5M aqueous sodium acetate for 2 hours, then the tetrahydrofuran isremoved in vacuo. The aqueous residue is acidified with dilutehydrochloric acid and extracted twice with ether. The ether extracts arecombined, dried and evaporated to an oil.

A mixture of this oil in 30 ml of chloroform and 60 ml of a solution of50 g of trimethylamine in 100 ml of acetonitrile is stirred at refluxfor 5 hours and then cooled overnight. The solvent is removed and theresidual oil stirred with 160 mg of silver carbonate and 30 ml ofmethanol for 2 hours, then filtered. The filtrate is evaporated and theresidue purified by chromatography, to give 267 mg of the desiredcompound as a glass.

EXAMPLE 63 Hexadecyl methyl-2-(phenylmethoxy)phosphoric acid, ethylester

To a solution of 28.4 1 of phosphorous oxychloride in 200 ml of carbontetrachloride is added dropwise a solution of 50 g of 1-hexadecanol in125 ml of carbon tetrachloride. This mixture is stirred overnight, andthe solvent removed. The residue is evaporated twice from toluene invacuo to give the phosphonyl dichloride as a dark oil.

A mixture of 156.75 g of dry ethylene glycol and 64.37 g of 87.2%potassium hydroxide is warmed and stirred until a complete solution. Themixture is then stirred in a 90° C. oil bath and 130.51 g of benzylchloride is added dropwise over 2 hours. This mixture is then stirred at130° C. for 2 hours, cooled and 500 ml of water added. The oil isextracted twice with ether. The extracts are combined and evaporated.The residue is distilled, giving 83.2 g of ethylene glycol monobenzylether.

To a solution of 34.4 g of phosphorodichloridic acid, hexadecyl ester in150 ml of carbon tetrachloride, stirred in an ice bath, is addeddropwise, at a fast rate, 27 ml of triethylamine. A solution of 10 g ofethylene glycol monobenzyl ether in 50 ml of carbon tetrachloride isadded dropwise. After 5 minutes, the mixture is removed from the icebath and stirred for 2 hours. A 100 ml portion of dry toluene is addedand the mixture filtered and washed with toluene. The filtrate and washare combined and evaporated. A 50 g portion of the residue is dissolvedin 350 ml of methanol and treated with 29 ml of triethylamine for 1hour, then evaporated. The residue is purified by chromatography, togive 16.2 g of the desired compound as a pale yellow oil.

EXAMPLE 64 Hexadecyl 2-hydroxyethyl phosphoric acid, methyl ester

A solution of 11.2 g of hexadecyl methyl 2-(phenylmethoxy)phosphoricacid, ethyl ester in 50 ml of glacial acetic acid and 100 ml of methanolis hydrogenated over 1.0 g of 10% palladium-on-carbon in a Parrapparatus at an initial pressure of 30 psi for 5 hours. Hydrogenation isrepeated overnight. The mixture is filtered, the filtrate evaporated andthe residue evaporated twice from toluene, to give 7.1 g of the desiredcompound as a waxy solid.

EXAMPLE 65 2-[[(2-Bromoethoxy)hydroxyphosphinyl]oxy]ethyl hexadecylphosphoric acid, methyl ester

To a solution of 4.52 g of 2-bromoethyl phosphorochloridate in 190 ml ofdry carbon tetrachloride, stirred in an ice bath under argon, is addeddropwise, 2.7 ml of triethylamine and a solution of 7.1 g ofhexadecyl-2-hydroxyethyl phosphoric acid, methyl ester in 30 ml of drycarbon tetrachloride. This mixture is stirred for 5 minutes, then theice bath is removed and stirring continued at ambient temperature for 2hours. A 100 ml portion of dry toluene is added and the mixture isfiltered and washed with toluene. The combined filtrate and wash isevaporated. The residue is hydrolyzed in a mixture of 150 ml oftetrahydrofuran and 150 ml of 0.5M aqueous sodium acetate with stirringfor 2 hours. The tetrahydrofuran is removed and the aqueous phaseacidified with dilute hydrochloric acid and extracted twice with ether.The extracts are combined, washed with saturated aqueous sodiumchloride, dried, filtered and evaporated. The residue is purified bychromatography, to give 5.17 g of the desired compound.

EXAMPLE 663,9-Dihydroxy-N,N,N-trimethyl-3,5,8,10-tetraoxa-4,9-diphosphahexacosan-1-aminium,hydroxide, 4-(inner salt) 4,9-dioxide

To a solution of 5.0 g of 2-[[(2-bromoethoxy)hydroxyphosphinyl]oxy]ethylhexadecyl phosphoric acid, methyl ester in 100 ml of dry acetonitrileand 90 ml of dry chloroform is added 50 g of anhydrous trimethylamine.This solution is stirred at reflux for 4 hours, then the solvent isremoved. The residue is stirred for 2 hours in 50 ml of methanolcontaining 1.215 g of silver carbonate, then filtered and washed withmethanol. The combined filtrate and wash is taken to dryness. Theresidue is purified by chromatography (65:43:10 chloroform:methylalcohol:water), to give 2.6 g of the desired compound.

EXAMPLE 67 Methyl octadecyl 2-(phenylmethoxy)phosphoric acid, ethylester

To a solution of 36.3 g of phosphoryl dichloride in 130 ml of carbontetrachloride, while stirring in an ice bath is added dropwise, at afast rate, 25.7 ml of triethylamine. A solution of 10 g ofphosphorodichloridic acid, octadecyl ester in 50 ml of carbontetrachloride is added dropwise. This mixture is stirred for 5 minutes,then the ice bath is removed and stirring continued at room temperaturefor 2 hours. A 100 ml portion of toluene is added, the mixture filteredand washed with toluene. The combined filtrates and wash arerefrigerated then evaporated. The residue is dissolved in 350 ml ofmethanol, 28 ml of triethylamine added and this solution stirred underargon for 1 hour. The solvent is removed, the residue dissolved in ethylacetate:hexane (1:1) and then evaporated to give an oil. The oil ispurified by chromatography, to give 25.8 g of the desired compound as awaxy solid.

EXAMPLE 68 2-Hydroxyethyl methyl phosphoric acid, octadecyl ester

A solution of 10.0 g of methyl octadecyl 2-(phenylmethoxy)phosphoricacid, ethyl ester in 100 ml of methanol and 50 ml of glacial acetic acidis hydrogenated over 1.0 g of 10% palladium-on-carbon at an initialpressure of 30 psi for 24 hours. The mixture is filtered and washed withmethanol. The combined filtrate and wash is evaporated and thenevaporated from toluene three times. The residue is purified bychromatography, to give 3.26 g of the desired compound.

EXAMPLE 694,9-Dihydroxy-N,N,N-trimethyl-3,5,8,10-tetraoxa-4,9-diphosphaoctacosan-1-aminium,hydroxide, 4-(inner salt), 4,9-dioxide

To a solution of 2.0 g of 2-hydroxyethyl methyl phosphoric acid,octadecyl ester in 75 ml of dry benzene, cooled in an ice bath underargon, is added with stirring 0.68 ml of triethylamine followed by asolution of 0.697 g of phosphoryl chloride in 5 ml of dry benzene. Thismixture is stirred for 5 minutes, then the ice bath removed and stirringcontinued for 24 hours. The solvent is removed leaving a white amorphoussolid.

This solid is slurried in 50 ml of dry acetonitrile, 10 ml of 33%trimethylamine in acetonitrile is added and this solution is transferredto a pressure bottle, rinsing with 5 ml of the trimethylamine solution.The bottle is sealed and the solution stirred at 65° C. overnight. Themixture is cooled and the insoluble material decanted, evaporated andthe two solids combined. This solid is purified by chromatography, togive 0.21 g of the desired compound.

EXAMPLE 70 Phosphorodichloridic acid, hexadecyl ester

To a moisture protected solution of 28.4 ml of phosphorous oxychloridein 200 ml of carbon tetrachloride is added dropwise with stirring asolution of 50 g of 1-hexadecanol in 125 ml of carbon tetrachloride.After stirring at ambient temperature for 18 hours, the solvent isremoved and the residue evaporated with toluene several times to give 71g of the desired product as a vacuum dried water white oil.

EXAMPLE 71 Hexadecyl 2-(hexadecyloxy)-1-[[(4-methoxyphenyl)diphenylmethoxy]methyl]ethyl phosphoric acid, methyl ester

To a solution of 7.5 g of1-[(4-methoxyphenoxy)diphenylmethoxy]-3-hexadecyloxy-2-propanol in 40 mlof dry carbon tetrachloride is added 1.0 g of dimethylaminopyridine and5.32 ml of triethylamine. A solution of 6.4 g of phosphorodichloridicacid, hexadecyl ester in 10 ml of dry carbon tetrachloride is addeddropwise at a fast rate with water bath cooling. This mixture is stirredat room temperature overnight, then 4.57 g of phosphorodichloridic acid,hexadecyl ester and 1.8 ml of triethylamine are added and stirringcontinued for 48 hours. An 8.5 ml portion of triethylamine and 250 ml ofmethanol are added, the mixture is stirred for 2 hours and then dilutedwith 100 ml of sodium bicarbonate solution. This mixture is extractedthree times with chloroform, the combined extracts dried and evaporatedand the residue purified by chromatography, to give 6.44 g of thedesired compound.

EXAMPLE 72 Hexadecyl 2-(hexadecyloxy)-1-(hydroxymethyl)ethyl phosphoricacid, methyl ester

A solution of 630 mg of hexadecyl2-(hexadecyloxy)-1-[[(4-methoxyphenyl)diphenylmethoxy]methyl]ethylphosphoric acid, methyl ester in 3 ml of methanol and 1.5 ml ofchloroform is heated to boiling. The mixture is removed from the heatand 300 mg of ion exchange resin added. This mixture is stirred for 1hour and 40 minutes, then filtered, diluted with chloroform, washed withaqueous sodium bicarbonate, dried and the solvent removed. The residueis chromatographed, to give 110 mg of the upper (less polar) isomer and120 mg of the lower (more polar) isomer.

EXAMPLE 73 3,5,8,10-Tetraoxa-4,9-diphosphahexacosan-1-aminium,7-[(hexadecyloxy)methyl]-4-hydroxy-9-methoxy-N,N,N-trimethyl- hydroxide,inner salt, 4,9-dioxide

To a solution of 1.6 g of methyl hexadecyl2-(hexadecyloxy)-1-(hydroxymethyl)ethyl phosphoric acid in 40 ml ofcarbon tetrachloride under an inert atmosphere is added 0.79 g ofphosphonyl dichloride followed by 0.5 ml of triethylamine. The mixtureis stirred at ambient temperature for 3 hours then stored in arefrigerator for 18 hours. After reaching ambient temperature, themixture is filtered and the cake washed with toluene. The volatiles areremoved and the residue dissolved in 50 ml of tetrahydrofuran and 50 mlof 0.5M sodium acetate. After stirring at ambient temperature for 2hours, the volatiles are removed and the residue acidified with dilutehydrochloric acid then extracted with ether. The organic layer is washedwith brine and dried. The volatiles are removed to give 1.6 g of a waxwhich is dissolved in 45 ml of chloroform, 50 ml of acetonitrile and 25g of trimethylamine. The mixture is stirred in an oil bath of 70° C. for4 hours. The volatiles are removed and the residue dissolved in 20 ml ofchloroform, 20 ml of methyl alcohol and 2 ml of water. While stirring,0.35 g of silver carbonate is rapidly added followed by continuedstirring at ambient temperature for 17 hours. The mixture is filteredthrough diatomaceous earth and the cake washed with 1:1 methylalcohol:chloroform. The volatiles are removed and the residuechromatographed on silica gel with 30% methyl alcohol:chloroformfollowed by 65:35:6 chloroform:methyl alcohol:water. The volatiles areremoved and the residue dissolved in ether, cooled and filtered to give310 mg of the desired product as a white solid.

EXAMPLE 74 3,5,8,10-Tetraoxa-4,9-diphosphahexacosan-1-aminium,7-[(hexadecyloxy)methyl]-4-hydroxy-9-methoxy-N,N,N-trimethyl-,hydroxide, inner salt, 4,9-dioxide

To a solution of 1.22 g of methyl hexadecyl2-(hexadecyloxy)-1-(hydroxymethyl)ethyl phosphoric acid in 30 ml ofcarbon tetrachloride under an inert atmosphere is added 0.6 g ofphosphonyl dichloride followed by 0.4 ml of triethylamine. The mixtureis stirred at ambient temperature for 5 hours then filtered throughdiatomaceous earth. The cake is washed with toluene and the combinedfiltrates evaporated to a syrup which is stirred at ambient temperaturefor 1.5 hours with 40 ml of tetrahydrofuran and 40 ml of 0.5M sodiumacetate. The tetrahydrofuran is evaporated and the aqueous residueacidified with dilute hydrochloric acid followed by ether extraction.The organic layer is washed with brine, dried and the solvent removed togive 1.6 g of a wax. The wax is dissolved in 35 ml of chloroform, 40 mlof a solution of 50 g of trimethylamine in 100 ml of acetonitrile andheated in an oil bath of 70° C. for 4 hours. The volatiles are removedand the residue stirred in a solution of 20 ml of chloroform, 20 ml ofmethyl alcohol and 2 ml of water containing 265 mg of silver carbonateat ambient temperature for 2 hours. The mixture is filtered throughdiatomaceous earth and the cake washed with 1:1 chloroform:methylalcohol. The volatiles are removed and the residue columned on silicagel with 30% methyl alcohol-chloroform followed by 65:35:6chloroform:methyl alcohol:water to give 287 mg of white wax aftersolvent evaporation and cooling the residue in ether.

EXAMPLE 75 3,5,8,10-Tetraoxa-4,9-diphosphahexacosan-1-aminium,7-[(hexadecyloxy)methyl]-4,9-dihydroxy-N,N,N-trimethyl-, hydroxide,4-inner salt), 4,9-dioxide

To a stirred solution of 2.38 g of phosphoric acid, hexadecyl2-(hexadecyloxy)-1-(hydroxymethyl)ethyl methyl ester, in 60 ml of carbontetrachloride is added 1.17 g of phosphoryl dichloride and by 0.73 ml oftriethylamine followed by stirring at ambient temperature for 3 hoursand allowing to stand in a refrigerator for 18 hours. After reachingambient temperature, the mixture is filtered and the cake washed withtoluene. The filtrate is taken to dryness and the residue stirred atambient temperature with a solution of 75 ml of tetrahydrofuran and 75ml of 0.5M sodium acetate for 2 hours. The tetrahydrofuran is removedand the aqueous residue acidified with dilute hydrochloric acid andextracted with ether. The separated organic layer is washed with brine,dried and evaporated to a syrup which is stirred in a solution of 45 mlof chloroform and 50 ml of acetonitrile containing 25 g oftrimethylamine at an oil bath temperature of 70° C. for 18 hours. Thevolatiles are removed and the residue stirred with 20 ml of chloroform,20 ml of methyl alcohol and 2.85 mg of silver carbonate at ambienttemperature for 2 hours. The mixture is filtered through diatomaceousearth and the cake washed with 1:1 chloroform:methyl alcohol. Thevolatiles are removed and the residue columned on silica gel with 30%methyl alcohol-chloroform and 65:35:6 chloroformmethyl alcohol-water.Fractions containing product are combined and evaporated. The residue isdissolved in ether and cooled in a refrigerator to give 560 mg of thedesired product as a white solid.

EXAMPLE 76 Phosphorodichloridic acid, pentyl ester

To a tap water cooled solution of 79 g of phosphorous oxychloride in 200ml of carbon tetrachloride is added dropwise with stirring a solution of50 g of 1-pentanol in 125 ml of carbon tetrachloride. The temperaturerises to 30° C. and stirring is continued for 18 hours. The solvent isevaporated and the syrup evaporated with toluene several times. Theresidue is distilled from a Kugelrohr still to give 108.7 g of thedesired product as a colorless oil, B.P. 85°-90° C./0.3 mm.

EXAMPLE 77 Phosphoric acid,2-(hexadecyloxy)-1-[[(4-methoxyphenyl)-diphenylmethoxy]methyl]ethylmethyl pentyl ester

To a stirred ice bath cooled solution of 15 g of1-[(4-methoxyphenoxy)diphenylmethoxy]-3-hexadecyloxy-2-propanol in 50 mlof carbon tetrachloride is added 2.1 g of dimethylaminopyridine followedby the addition of 8.2 ml of triethylamine. A solution of 7.31 g ofphosphorodichloridic acid, pentyl ester in 10 ml of carbon tetrachlorideis added dropwise, the cooling bath removed and the reaction stirred atambient temperature for 66 hours. A solution of 10 ml of triethylamineand 500 ml of methyl alcohol is added followed by continued stirring for2 hours. The reaction is diluted with 1500 ml of 5% sodium bicarbonateand extracted with chloroform. The organic layer is dried and thevolatiles removed to give 21 g of an oil which is dry columned on silicagel with 1:4 ethyl acetate:hexanes. Product fractions are combined inchloroform and the solvent removed to give 10 g of the desired productas a yellow oil.

EXAMPLE 78 Phosphoric acid, 2-(hexadecyloxy)-1-(hydroxymethyl) ethylmethyl pentyl ester, more polar isomer Phosphoric acid,2-(hexadecyloxy)-1-(hydroxymethyl) ethyl methyl pentyl ester, less polarisomer

A solution of 10 g of phosphoric acid,2-(hexadecyloxy)-1-[[(4-methoxyphenyl)-diphenylmethoxy]methylethylmethyl pentyl ester in 100 ml of methyl alcohol and 50 ml of chloroformis brought to a boil, the heat removed and 5 g of Amberlite®-15 resincarefully added. The reaction is stirred at ambient temperature for 2hours, filtered to remove the resin and the cake washed well with 2:1methyl alcohol:chloroform. The solvent is removed to give 9.7 g of athick syrup. The product is chromatographed using dry column silica gelwith 1:1 ethyl acetate:hexanes. Product fractions are combined and thesolvents evaporated to give 1.38 g of a less polar isomer oily productand 1.46 g of a more polar isomer oily product.

EXAMPLE 79 2-Bromoethyl Phosphorodichloridate

To a stirred solution of 412.98 g of phosphorous oxychloride in 910 mlof carbon tetrachloride is added 224.4 g of 2-bromoethanol with stirringover 30 minutes. The temperature increases to 34° C. and the reaction isstirred under a dry atmosphere for 18 hours. The solvent is removed to asyrup which is evaporated with toluene several times. The residue isdistilled to afford 161 g of the desired product as a colorless oil,B.P. 82° C./0.3 mm.

EXAMPLE 80 3,5,8,10-Tetraoxa-4,9-diphosphapentadecan-1-aminium,7-[(hexadecyloxy)methyl]-4-hydroxy-9-methoxy-N,N,N-trimethyl-,hydroxide, 4-(inner salt),4,9-dioxide, more polar isomer3,5,8,10-Tetraoxa-4,9-diphosphapentadecan-1-aminium,7-[(hexadecyloxy)methyl]-4,9-dihydroxy-N,N,N-trimethyl-, hydroxide,4-(inner salt), 4,9-dioxide, less polar isomer

To a stirred solution of 1.46 g of phosphoric acid,2-(hexadecyloxy)-1-(hydroxymethyl)ethyl methyl pentyl ester, lowerdiastereomer in 35 ml of carbon tetrachloride, cooled in an ice bathunder argon, is added dropwise a solution of 1.0 g of 2-bromoethylphosphorodichloridate in 1 ml of carbon tetrachloride followed by 0.7 mlof triethylamine. The bath is removed and the mixture stirred at ambienttemperature for 1 hour followed by storing in a refrigerator for 18hours. The mixture is brought to ambient temperature and filteredthrough diatomaceous earth. The cake is washed with toluene and thecombined solvents evaporated. The residue is hydrolyzed by stirring for2 hours at ambient temperature with 50 ml of tetrahydrofuran and 50 mlof 0.5M sodium acetate. The tetrahydrofuran is evaporated and theaqueous residue acidified with hydrochloric acid followed by etherextraction. The organic layer is washed with brine, dried and thesolvent removed to give 1.8 g of a stiff oil which is dissolved in 40 mlof chloroform and 40 ml of a solution of 50 g of trimethylamine inacetonitrile then heated in an oil bath of 65° C. for 4 hours. Thesolvent is evaporated and the residue dissolved in 30 ml of methylalcohol followed by stirring with 450 mg of silver carbonate at ambienttemperature for 1.5 hours. The mixture is filtered and the filtrateevaporated to a residue which is chromatographed on silica gel andeluted with 30% methyl alcohol:chloroform, and then 65:35:6chloroform:methyl alcohol:water. The solvent is evaporated to give 253mg of the less polar isomer product as a glass. The column is furthereluted with the same solvent system to give 275 mg of the more polarisomer product as a white solid.

EXAMPLE 8 3,5,8,10-Tetraoxa-4,9-diphosphapentadecan-1-aminium,7-[(hexadecyloxy)methyl]-4-hydroxy-9-methoxy-N,N,N-trimethyl-,hydroxide, 4-(inner salt), 4,9-dioxide, less polar isomer

To a solution of 1.28 g of phosphoric acid,2-(hexadecyloxy)-1-(hydroxymethyl)ethyl methyl pentyl ester, upperdiastereomer in 35 ml of carbon tetrachloride, cooled in an ice bathunder an inert atmosphere is added rapidly dropwise 0.9 g of2-bromoethyl phosphorodichloridate followed by the rapid dropwiseaddition of 0.6 ml of triethylamine. The bath is removed and the mixturestirred at ambient temperature for 1 hour then stored in therefrigerator for 18 hours. After reaching room temperature, the mixtureis filtered through diatomaceous earth rinsing the cake with toluene.The solvents are removed and the residue stirred at room temperaturewith 50 ml of tetrahydrofuran and 50 ml of 0.5M sodium acetate for 2hours. The tetrahydrofuran is removed and the aqueous residue acidifiedwith hydrochloric acid and extracted with ether. The organic layer iswashed with brine, dried and evaporated to give 1.9 g of a stiff oil,which is dissolved in 40 ml of chloroform and 40 ml of a solutioncontaining 50 g of trimethylamine in 100 ml of acetonitrile. The mixtureis heated in an oil bath of 65° C. for 4 hours. The solvent is removedand the residue stirred in 30 ml of methyl alcohol containing 400 mg ofsilver carbonate at ambient temperature for 1.5 hours. The mixture isfiltered through diatomaceous earth and the filtrate evaporated to give2 g of a glass which is chromatographed on silica gel with 30% methylalcohol:chloroform and 65:35:6 chloroform:methyl alcohol:water.Fractions containing product are combined and the volatiles removed togive 300 mg of the desired product as a dried glass.

EXAMPLE 827-[(Chloroacetyl)oxy]-4-hydroxy-N,N,N-trimethyl-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide

A mixture of 500 mg of4-hydroxy-N,N,N-tri-methyl-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 86.2 mg of anhydrous sodium acetate and 1.8 g ofchloroacetic anhydride is stirred at reflux under argon in 25 ml ofchloroform for 2 hours then stirred at room temperature for four days.An additional 500 mg of chloroacetic anhydride is added and the mixturestirred at reflux under argon for 5 hours. The solvent was removed invacuo and the concentrate columned on silica gel with 30%methanol-chloroform. Further elution with 65:35:6chloroform:methanol:water affords 155 mg of the desired product as aglass following solvent removal and drying in vacuum.

EXAMPLE 8310-Bromo-7-[(hexadecyloxy)methyl]-4-hydroxy-N,N,N-trimethyl-9-oxo-3,5,8-trioxa-4-phosphahexacosan-1-aminium,hydroxide, inner salt, 4-oxide

A solution of 455 mg of4-hydroxy-N,N,N-trimethyl-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide and 472 mg of 2-bromostearic acid in 15ml of dichloromethane is stirred at room temperature under argon while402 mg of dicyclohexylcarbodiimide is rapidly added followed by 23.8 mgof dimethylaminopyridine. Stirring is continued for 18 hours. Themixture is filtered and the solvent removed in vacuo to give 1.1 g of aglass. The product is chromatographed on silica gel using 30%methanol-chloroform. Further elution with 65:35:6chloroform:methanol:water affords 422 mg of a glass which is trituratedwith ether and cooled to afford 332 mg of the desired product as a whiteamorphorous solid.

EXAMPLE 84 3,5,9-Trioxa-4-phosphapentacosan 1-aminium,7-[(bromoacetyl)oxy]-4-hydroxy-N,N,N-trimethyl-, hydroxide, inner salt,4-oxide

A mixture of 500 mg of 3,5,9-trioxa-4-phosphapentacosan-1-aminium,4-hydroxy, N,N,N-trimethyl, hydroxide, inner salt, 4-oxide, 86.2 mg ofsodium acetate and 1.35 g of bromoacetic anhydride in 25 ml ofchloroform is stirred at an oil bath of 70° C. for 5 hours then allowedto cool overnight. The solvent is removed and the residue columned onsilica gel with 30% methyl alcohol:chloroform followed by 65:35:6chloroform:methyl alcohol:water to give 198 mg of the desired product asa glass.

EXAMPLE 854-[2-(Acetyloxy)-3-(octadecyloxy)propoxy]-N,N,N-trimethyl-1-butanaminiumbromide

A solution of 580 mg of4-[2-hydroxy-3-(octadecyloxy)-propoxy]-N,N,N-trimethyl-1-butanaminiumbromide in 5 ml of acetic anhydride is stirred at reflux under inert gasfor 15 minutes. Toluene is added and the solvents evaporated followed byseveral additional toluene evaporations. The residue is stirred withether and stored at 0° C. for several days. The resulting solid iscollected and dried to give 480 mg of the desired product.

EXAMPLE 86 3-(Pentyloxy)-1,2-propanediol

To a mixture of 18.6 g of hexane washed sodium hydride in 400 ml oftoluene under inert gas is added dropwise over 1 hour a solution of 60 gof solketal in 100 ml of toluene. Stirring is continued for 30 minutesat ambient temperature and a solution of 57 g of 1-bromopentane in 100ml of toluene is added dropwise. The mixture is heated at reflux for 1.5hours and then allowed to cool at ambient temperature for 18 hours. A500 ml volume of ice water is carefully added. The organic layer isseparated, washed with saturated sodium chloride and dried. The solventis removed and the residual oil stirred at reflux with 900 ml of methylalcohol and 100 ml of 1N hydrochloric acid for 2 hours. The solvent isremoved and the syrup dissolved in chloroform. The aqueous layer isremoved and the organic layer dried and evaporated. The concentrate isvacuum distilled in a Kugelrohr apparatus to give 49.4 g (bp 85°-86°C./0.15 mm) of the desired product as a colorless oil.

EXAMPLE 87 1[(4-methoxyphenoxy)diphenylmethoxy]-3-(pentyloxy)-2-propanol

To an ice bath cooled solution of 48 g of 3-(pentyloxy)-1,2-propanediolin 150 ml of dry pyridine under inert gas is added at a rapid rate awarm solution of 127.92 g of p-methoxytrityl chloride in 165 ml oftetrahydrofuran. The mixture is stirred for an additional 20 minutes atice bath temperature followed by 2 hours at ambient temperature and 18hours in a refrigerator. The reaction mixture is filtered and the cakewashed with ether. The combined filtrate and washings are evaporated toa residue which is dissolved in chloroform and washed with 10% sodiumbicarbonate, water and dried to give a thick oil upon evaporation of thevolatiles. The residue is purified by high pressure liquidchromatography on silica gel using 1:9 ethyl acetate:hexanes to give 96g of the desired product as an oil.

EXAMPLE 88 Phosphoric Acid.1-(4-methoxyphenyl)diphenylmethoxy]methyl]-2-(pentyloxy)ethyl methylpentyl ester

To a solution of 23.8 g of1-[(4-methoxyphenoxy)diphenylmethoxy]-3-(pentyloxy)-2-propanol in 80 mlof dry carbon tetrachloride under inert gas is added 4.54 g ofdimethylaminopyridine followed by 35.1 ml of triethylamine. Whilecooling in an ice bath, a solution of 31.44 g of phosphorodichloridicacid, pentyl ester in 45 ml of carbon tetrachloride is added dropwise ata moderate rate. The water bath is removed and stirring continued for 4days. Added 16 ml of triethylamine and 790 ml of methyl alcohol followedby stirring at ambient temperature for 2 hours. Diluted with 2350 ml of5% sodium bicarbonate and extracted with chloroform. The extract isdried and evaporated to an oil which is purified on silica gel with30:70 ethyl acetate:hexanes to give 15.1 g of the desired product as ayellow thick oil.

EXAMPLE 89 Phosphoric acid, 1-(hydroxymethyl)-2 (pentyloxy)ethyl methylpentyl ester

To a solution of 13.5 g of phosphoric acid,1-[[(4-methoxyphenyl)diphenylmethoxy]-methyl]-2-(pentyloxy)ethyl methylpentyl ester in 68 ml of chloroform and 136 ml of methyl alcohol heatedto boiling is slowly added 6.8 g of Amberlyst®-15 resin. The heat isremoved and the mixture stirred at ambient temperature for 2 hours. Thevolatiles are removed and the residue dissolved in chloroform thenwashed with dilute sodium bicarbonate, dried and evaporated to an oilyresidue which is purified on silica gel using 1:1 ethyl acetate:hexanesto give 6 g of the desired compound as a colorless oil.

EXAMPLE 90 Phosphoric acid,1-[[[(2-bromoethoxy]hydroxyphosphinyl]oxy]1methyl]-2-(pentyloxy]ethylmethyl pentyl ester

To a stirred solution of 6.0 g of phosphoric acid1-(hydroxymethyl)-2-(pentyloxy)ethyl methyl pentyl ester in 140 ml ofcarbon tetrachloride while cooling in an ice bath under inert gas isadded very rapidly a solution of 5.8 g of 2-bromoethylphosphorodichloridate in 6 ml of carbon tetrachloride. The ice bath isremoved and stirring continued for 3 hours at ambient temperature. Added100 ml of toluene, stirred for a short time and filtered. The cake iswashed with additional toluene and the filtrates are evaporated to aresidue which is stirred at ambient temperature for 1.5 hours with 125ml of 0.5M sodium acetate and 125 ml of tetrahydrofuran. Thetetrahydrofuran is removed to an aqueous residue which is cooled andacidified with hydrochloric acid until a gum separates. The etherextracts are dried and evaporated to a residue which is evaporated withtoluene several times, then dried with a vacuum pump. The residue ispurified on 100 g of magnesium silicate with chloroform followed by 10%methyl alcohol:chloroform. Fractions containing the desired product areevaporated to give 5.6 g of the desired product as a stiff oil.

EXAMPLE 91 3,5,8,10-Tetraoxa-4,9 diphosphapentadecan-1-aminium,4-hydroxy-9-methoxy-N,N,N-trimethyl-7-[(pentyloxy) methyl]-, hydroxide,inner salt, 4,9-dioxide, more polar isomer3,5,8,10-Tetraoxa-4,9-diphosphapentadecan-1-aminium,4,9-dihydroxy-N,N,N-trimethyl-7-[(pentyloxy)methyl]-, hydroxide,4-(inner salt), 4,9-dioxide, less polar isomer

To 5.2 g of phosphoric acid,1-[[[(2-bromoethoxy)hydroxyphosphinyl]oxy]methyl]-2-(pentyloxy)ethylmethyl pentyl ester in 115 ml of dry chloroform is added 130 ml of dryacetonitrile and 50 ml of anhydrous trimethylamine. The solution isheated at 62° C. for 4 hours and the solvents removed. The residue isdissolved in 70 ml of methyl alcohol and stirred with 1.4 g of silvercarbonate at ambient temperature for 2 hours. The reaction mixture isfiltered through diatomaceous earth followed by washing with methylalcohol. The combined filtrates are evaporated to a glass which ispurified by chromatography on silica gel eluting with 30% methylalcohol:chloroform then chloroform:methyl alcohol:water, 65:35:6 to give395 mg of the less polar isomer product as a glass. Further elution ofthe column gives 2.7 g of the second more polar isomer product as aglass.

EXAMPLE 92 (R)-4-](Hexadecyloxy)methyl]-2,2-dimethyl-1,3-dioxolane

To a mixture of 3.2 g of hexanes washed sodium hydride in 67 ml of drytoluene while stirring under inert gas is added dropwise over 1 hour asolution of 10.52 g of L-2,3-O-isopropylidene-sn-glycerol in 17 ml ofdry toluene. Following 15 minutes of stirring, a solution of 24.1 g ofhexadecylbromide in 17 ml of toluene is added dropwise over 20 minutes.The mixture is gradually heated to reflux followed by 1.5 hours atreflux and 18 hours at ambient temperature. The reaction mixture isfiltered, the cake washed with toluene and the combined filtrates washedwith water and brine then dried. The reaction mixture is filtered andthe solvent removed to give 28.2 g of the desired product as a yellowoil.

EXAMPLE 93 (S)-3-(Hexadecyloxy)-1,2-propanediol

A mixture of 28.2 g of(R)-4-[(hexadecyloxy)methyl]-2,2-dimethyl-1,3-dioxolane in 189 ml ofmethyl alcohol and 21 ml of 1N hydrochloric acid is stirred at refluxfor 2.5 hours followed by 12 hours at ambient temperature. The reactionis cooled, filtered and the cake washed with cold methyl alcohol thendried to give 20.3 g of the desired product as white plates, m.p.63°-64° C.

EXAMPLE 94 (R)-1-(Hexadecyloxy)-3-[(4-methoxyphenyl)diphenylmethoxy]-2-propanol

To a solution of 15.34 g of (S)-3-(hexadecyloxy)-1,2-propanediol in 50ml of pyridine is added dropwise with stirring a solution of 20.96 g ofp-methoxytrityl chloride in 30 ml of tetrahydrofuran. Stirring iscontinued at ambient temperature for 2 hours followed by refrigerationfor 18 hours. The mixture is filtered and the cake washed with ether.The combined filtrates are evaporated to a residue which is dissolved inchloroform and washed with 10% sodium bicarbonate and water then driedand evaporated to an oil which is purified by high pressure liquidchromatography on silica gel using 1:9 ethyl acetate:hexanes to give28.44 g of the desired product as an oil.

EXAMPLE 95 Phosphoric acid, (R1-hexadecyl2-(hexadecyloxy)-1-[[(4-methoxyphenyl)diphenylmethoxy]methyl]ethylmethyl ester

To a stirred solution of 14 g of(R)-1-(hexadecyloxy)-3-[(4-methoxyphenyl)diphenylmethoxy]-2-propanol in60 ml of carbon tetrachloride under inert gas is added 2 g of4-dimethylaminopyridine and 14.24 ml of triethylamine. A solution of21.94 g of phosphorodichloridic added dropwise at a rapid rate. Stirringis continued at ambient temperature for 4 days. A solution of 17 ml oftriethylamine and 500 ml of methyl alcohol is added followed by stirringat ambient temperature for 2 hours. The reaction mixture is poured into1800 ml of 10% sodium bicarbonate and extracted several times withchloroform. The combined extracts are dried and evaporated to a residuewhich is purified by high pressure liquid chromatography on silica gelusing 1:7 ethyl acetate:hexanes to give 13 g of the desired product asan oil.

EXAMPLE 96 Phosphoric acid, hexadecyl2-(hexadecyloxy)-1-(hydroxymethyl)ethyl methyl ester, (1S)-, upperdiastereomer Phosphoric acid, hexadecyl2-(hexadecyloxy)-1-hydroxymethyl)ethyl methyl ester, (1S)-, lowerdiastereomer

To a solution of 13 g of phosphoric acid, (R)-hexadecyl2-(hexadecyloxy)-1-[[(4-methoxyphenyl) methoxy]methyl]ethyl methylester, in 90 ml of methyl alcohol and 45 ml of chloroform brought to agentle boil is added 7.7 g of Amberlyst®-15 resin. The mixture isstirred at ambient temperature for 1.5 hours then filtered. The filtrateis evaporated to a residue which is dissolved in chloroform, washed withdilute sodium bicarbonate and dried. The solvent is evaporated to aresidue which is purified by chromatography on silica gel using 1:1ethyl acetate:hexanes to afford 3.6 g of an upper diastereomer and 3.1 gof lower diastereomer.

EXAMPLE 97 3,5,8,10-Tetraoxa-4,9-diphosphahexacosan-1-aminium,7-[(hexadecyloxy)methyl]-4,9-dihydroxy-N,N,N-trimethylhydroxide,4-(inner salt), 4,9-dioxide, more polar isomer

To a solution of 3.1 g of phosphoric acid, hexadecyl2-(hexadecyloxy)-1-hydroxy-methyl)ethylmethyl ester, (1S)-, more polardiastereomer in 50 ml of carbon tetrachloride stirred in an ice bathunder an argon atmosphere is added rapidly 1.5 g of 2-bromoethylphosphorodichloridate in 0.5 ml of carbon tetrachloride followed by therapid addition of 0.95 ml of triethylamine. Stirring is continued atambient temperature for 1.5 hours followed by storing in a refrigeratorfor 18 hours. The reaction mixture is filtered through diatomaceousearth and the cake washed with toluene. The combined filtrates areevaporated to a residue which is stirred with 110 ml of 0.5M sodiumacetate and 110 ml of tetrahydrofuran for 1.5 hours. The tetrahydrofuranis evaporated and the aqueous residue acidified with 10% hydrochloricacid and extracted with ether. The ether is washed with brine, dried,then evaporated to a wax residue which is dissolved in 90 ml ofchloroform and 100 ml of a solution of 50 g of trimethylamine in 100 mlof acetonitrile added. The mixture is stirred at reflux for 4 hours. Thesolvents are evaporated and the residue dissolved in 50 ml of chloroformand 50 ml of methyl alcohol containing 5 ml of water and 673 mg ofsilver carbonate then stirred at ambient temperature for 2 hours. Themixture is filtered through diatomaceous earth, washing with 1:1 methylalcohol:chloroform. The solvents are evaporated to a glass which ispurified by chromatography on silica gel by eluting with 30% methylalcohol:chloroform followed by 65:35:6 chloroform:methyl alcohol:water.Fractions are combined and evaporated to a residue which is stirred withether and filtered to give 0.74 g of the less polar isomer product as anamorphous solid. Further elution of the column with the same solventmixture followed by evaporation of product fractions to a residue whichis stirred with ether and filtered to give 0.87 g of the more polarisomer product as a white solid. The more polar isomer product (0.74 g)is purified by chromatography on silica gel with 30% methylalcohol:chloroform followed by 65:35:6 chloroform:methyl alcohol:water.Fractions containing product are combined and evaporated to a residuewhich was stirred with ether and filtered to give 0.58 g of the desiredproduct as a white amorphous solid.[α]_(D) ²⁶ =4.5°±0.3.

EXAMPLE 98 3,5,8,10-Tetraoxa-4,9-diphosphahexacosan-1-aminium,(7R)-,7-[(hexadecyloxy)methyl]-4-hydroxy 9 methoxy-N,N,N-trimethyl-,hydroxide, inner salt, 4,9-dioxide, less polar diastereomer

To a stirred solution of 3.6 g of phosphoric acid, (1S)-hexadecyl2-(hexadecyloxy)-1-(hydroxymethyl)ethyl methyl ester, less polardiastereomer in 50 ml of dry carbon tetrachloride while cooling in anice bath under inert gas is added 1.8 g of 2-bromoethylphosphorodichloridate followed by the rapid dropwise addition of 1.1 mlof triethylamine. The cooling bath is removed and the reaction stirredat ambient temperature for 3 hours followed by storing in a refrigeratorfor 18 hours. The reaction mixture is filtered and the filter cakewashed with toluene. The combined filtrates are evaporated and theresidue stirred with 110 ml of 0.5M sodium acetate and 110 ml oftetrahydrofuran at ambient temperature for 1.5 hours. Thetetrahydrofuran is removed and the aqueous residue acidified with dilutehydrochloric acid followed by ether extraction. The ether extract iswashed with brine, dried and evaporated to give a waxy residue which isdissolved in 90 ml of chloroform to which 100 ml of a solution of 50 gof trimethylamine in 100 ml of dry acetonitrile is added. The mixture isrefluxed by heating in a 70° C. bath for 4 hours. The volatiles areremoved and the residue dissolved in 60 ml of chloroform, 60 ml ofmethyl alcohol and 6 ml of water to which 782 mg of silver carbonate isadded followed by stirring at ambient temperature for 2 hours. Themixture is filtered through diatomaceous earth and the cake washed with1:1 methyl alcohol:chloroform. The combined filtrates are evaporated toa residue which is purified by chromatography on silica gel using 30%methyl alcohol:chloroform followed by 65:35:6 chloroform: methylalcohol:water. Fractions containing the desired product are combined andevaporated to a residue which is stirred with ether and filtered to give646 mg of less polar isomer product. Further elution followed byevaporation of solvents affords 896 mg of a residue [α]_(D) ²⁶ +3.2°±0.6c=1.55% in chloroform solvent. The less polar isomer product is furtherpurified by chromatography on silica gel using 30% methylalcohol:chloroform followed by 65:35:6 chloroform:methyl alcohol: waterto give upon evaporation of solvents followed by additional evaporationwith toluene and stirring the residue with ether, 480 mg of the lesspolar isomer produce as a white powder. [α]_(D) ²⁶ +4.7°±2 c=4.235% inchloroform solvent.

EXAMPLE 99 3,5,9-Trioxa-4-phosphapentacosan-1-aminium,7-[(bromoacetyl)oxy]-4-hydroxy-N,N,N-trimethyl-, hydroxide, inner salt,4-oxide

A mixture of 500 mg of 3,5,9-trioxa-4-phosphapentacosan-1-aminium,4-hydroxy-N,N,N-trimethyl-, hydroxide, inner salt, 4-oxide, 86.2 mg ofanhydrous sodium acetate and 1.35 g of bromoacetic anhydride in 25 ml ofchloroform is stirred in an oil bath of 75° C. for 5 hours then allowedto cool over 18 hours. The chloroform is evaporated and the residuepurified by chromatography on silica gel using 30% methylalcohol:chloroform followed by chloroform:methyl alcohol:water to give198 mg of the desired product as a glass.

EXAMPLE 100 Ethyl 2-[(hexadecyloxy]methyl]-2-propenoate

To a stirred solution of 1.9 g of hexadecanol in 40 ml of methylenechloride containing 700 mg of dimethylaminopyridine is added 500 mg ofethyl 2-bromomethyl-2-propenoate followed by continued stirring for 18hours. The solvent is evaporated to give 2.4 g of the desired product.

EXAMPLE 101 Ethyl 3-(hexadecyloxy)-2-[(hexadecylthio)methyl]propanoate

To a stirred solution of 20 g of ethyl2-[(hexadecyloxy)methyl]-2-propenoate in 60 ml of ethyl alcoholcontaining 10 ml of triethylamine is added 53 ml hexadecylmercaptan.Following stirring for 18 hours, the volatile components are removed andthe residue is chromatographed by HPLC on silica gel using 9:1hexanes:ethyl acetate. Fractions containing product are combined and thesolvents evaporated to give 32.9 g of the desired product as a whitewaxy solid.

EXAMPLE 102 3-(Hexadecyloxy)-2-(hexadecylthio)methyl-1-propanol

To a slurry of 1.04 g of lithium borohydride in tetrahydrofuran atambient temperature is added 29 g of ethyl3-(hexadecyloxy)-2-[(hexadecylthio)methyl]propanoate followed by heatingat reflux for 18 hours. The mixture is poured into water containing asmall amount of acid followed by extraction with ether to give afterdrying and evaporation, a residue which is columned on silica gel using30:1 hexane:ethyl acetate. Fractions containing the product areevaporated to give 18.4 g of the desired product.

EXAMPLE 103 Phosphoric acid, 2-bromoethyl3-(hexadecyloxy)-2-[(hexadecylthio)methyl]propyl ester

A mixture of 15.0 g of3-(hexadecyloxy)-2-[(hexadecylthio)methyl]-1-propanol, 9.6 g ofphosphate and 5.7 ml of triethylamine in 100 ml of carbon tetrachlorideis stirred at room temperature for 3 hours. Following filtration throughdiatomaceous earth, the solvents are evaporated and the residue stirredwith 100 ml of 0.5M sodium acetate and 100 ml of tetrahydrofuran atambient temperature for 18 hours. Dilute hydrochloric acid is addedfollowed by extraction with ethyl acetate. The combined extracts aredried and evaporated to give 18.3 g of the desired product as a solid,m.p. 48°-50° C.

EXAMPLE 1047-[(Hexadecylthio)methyl]-4-hydroxy-N,N,N-trimethyl-3,5,9-trioxa-4-phosphapentacosan-1-aminium,hydroxide, inner salt, 4-oxide

A solution of 25.76 g of phosphoric acid,2-bromoethyl]3-(hexadecyloxy)-2-[(hexadecylthio)methyl]propyl ester in200 ml of acetonitrile, 150 ml of chloroform and 173 ml oftrimethylamine is heated at 60° C. for 18 hours. The solvents areevaporated and the residue is stirred with 150 ml of methyl alcohol, 4.0g of silver carbonate and 3.0 g of Amberlite®IR-4B resin for 2 hours.The insolubles are filtered and the filtrate evaporated to a residuewhich is chromatographed on silica gel using 70:30 chloroform:methylalcohol and 65:35:6 chloroform:methyl alcohol:water to give uponevaporation of fractions containing the product, a residue which ischromatographed on silica gel using high pressure liquid chromatographywith 65:35:6 chloroform:methyl alcohol:water to give 8.5 g of thedesired compound as a white solid.

EXAMPLE 105 Ethyl 2-[(hexadecyloxy)methyl]-2-propenoate

A solution of 400 mg of 1.1 g of hexadecanol and 550 mg ofdimethylaminopyridine in 15 ml of methylene chloride is stirred atambient temperature for 18 hours. The solvent is removed and the residuesubjected to preparative thick layer chromatography using 25% ethylacetate:petroleum ether to give 324 mg of the desired product as a gum.

EXAMPLE 106 2-[(Hexadecyloxy)methyl]-2-propen-1-ol

To a solution of 21 g of ethyl 2-[(hexadecyloxy)methyl]-2-propenoate in400 ml of ether cooled to 0° C. under an inert atmosphere is added 2.25ml of 1,2M DIBAL in hexanes over 30 minutes. The reaction is stirred atambient temperature for 2 hours then cooled to 0° C. while 20 ml ofsaturated sodium sulfate is added dropwise. After filtering, the cake iswashed with ether. The combined ether washings are dried and the solventremoved. The residue is crystallized from methyl alcohol:water to give25.8 g of the desired product as a colorless solid.

EXAMPLE 107 Ethaniminium,2-[[hydroxy[(2-methylenenonadecyl)oxy]phosphinyl]oxy]-N,N,N-trimethyl-,hydroxide, inner salt

To a solution of 3.8 g of 2-[(hexadecyloxy)methyl]-2-propen-1-ol in 100ml of carbon tetrachloride is added 3.91 g of 2-bromoethylphosphorodichloridate and 1.64 g of triethylamine followed by storing ina refrigerator for 18 hours. The reaction mixture is filtered and thesolvent removed. The residue is stirred with 100 ml of 0.5M sodiumacetate and 100 ml of tetrahydrofuran for 1 hour. The tetrahydrofuran isremoved and the residue acidified with dilute sulfuric acid followed byether extraction. The organic layer is washed with brine and dried. Thesolvent is removed and the residue stirred at reflux in 60 ml ofacetonitrile, 50 ml of chloroform and 20 g of trimethylamine for 4hours. The solvent is removed and the residue stirred with 100 ml ofmethyl alcohol, 10 g of Amberlite® IR-4B resin and 1 g of silvercarbonate then filtered. The solvent is evaporated and the residuechromatographed on silica gel eluting with 7:3 chloroform:methyl alcoholand then with 70:30:1 chloroform:methyl alcohol:water. The volatiles areremoved and the fractions containing the desired product triturated withether and filtered to give 2.4 g of the desired product as a whitesolid.

EXAMPLE 108 3,5,9-Trioxa-4-phosphapentacosan-1-aminium,7-(boronomethyl)-4-hydroxy-N,N,N-trimethyl-, hydroxide, inner salt,4-oxide

To a stirred solution of 9 ml of 0.98M Borane in tetrahydrofuran at 0°C. under inert gas is added dropwise over 20 minutes a solution of 2.4 gof ethaniminium,2-[[hydroxy[(2-methylenenonadecyl)oxy]phosphinyl]oxy]-N,N,N-trimethyl-,hydroxide, inner salt in 75 ml of methylene chloride. Stirring iscontinued for 1.5 hours. The volatiles are removed and methyl alcoholadded to the residue. The methyl alcohol is removed by evaporation andthe concentrate chromatographed on silica gel with 7:3 chloroform:methylalcohol followed by 70:30:3.5 chloroform:methyl alcohol:water to give0.72 g of the desired product as a white powder from ether trituration.

EXAMPLE 109 [2-(Hexadecyloxy)-1-(hydroxymethyl)-ethoxy]acetic acid

To a stirred slurry of 13.07 g of hexane washed sodium hydride in 200 mlof toluene is added dropwise a solution of 12.61 g of chloroacetic acidin 50 ml of toluene under an inert gas over 45 minutes. A solution of65.5 g of1-[(4-methoxyphenoxy)diphenylmethoxy]-3-hexadecyloxy-2-propanol in 100ml of toluene is added dropwise over 30 minutes followed by reflux for65 hours. The pH is adjusted to 4 with dilute hydrochloric acid and themixture extracted with ether. The organic layer is washed with brine,dried and the solvent removed. The residue is stirred for 1 hour with450 ml of methyl alcohol, 100 ml of chloroform and 20 g of Amberlite®IR-4B resin. The solvents are evaporated and the residue chromatographedon silica gel by high pressure liquid chromatography using 4:1hexane:ethyl acetate. Fractions containing product are evaporated togive 22.39 g of the desired product as a white solid.

EXAMPLE 110 2,5,7-Trioxa-6-phosphanonan-9-aminium,1-carboxy-3-[(hexadecyloxy)methyl]-6-hydroxy-N,N,N-trimethyl-,hydroxide, inner salt, 6-oxide

To a stirred solution of 22.39 g of[2-(hexadecyloxy)-1-(hydroxymethyl)-ethoxy]acetic acid in 150 ml ofcarbon tetrachloride is added 6.48 g of triethylamine and 11.62 g of2-bromoethyl phosphorodichloridate. The mixture is stirred for 1.5 hoursand filtered. The solvent is removed and the residue was stirred for 2hours with 300 ml of 0.5M sodium acetate and 300 ml of tetrahydrofuranfollowed by acidification with hydrochloric acid and ether extraction.The ether extract is dried and evaporated to a residue which is refluxedwith 250 ml of acetonitrile, 180 ml of chloroform, 80 g oftrimethylamine and 20 g of Amberlite® IR-4B resin for 4 hours. Thesolvent is removed and the residue chromatographed on silica gel with9:1 chloroform:methyl alcohol followed by 70:30:3 chloroform:methylalcohol:water to give 9.07 g of the desired product as a white solidafter stirring the fraction concentrates with ether and filtering.

EXAMPLE 111 Ethyl 3-(hexadecylthio)propanoate

A solution of 15 g of hexadecylmercaptan, 3.0 g of ethyl acrylate and 2ml of triethylamine is stirred at ambient temperature for 18 hours. Thesolvent is evaporated and the concentrate purified by high pressureliquid chromatography, to give 19.8 g of the desired product.

EXAMPLE 112 3-(Hexadecylthio)-1-propanol

To a slurry of 147 mg of lithium borohydride in 20 ml of tetrahydrofuranis added slowly with stirring 2.4 g of ethyl3-(hexadecylthio)propanoate. The mixture is refluxed for 18 hours andpoured into water. A few drops of hydrochloric acid is added followed byether extraction. The organic layer is dried and evaporated to give 2.1g of the desired product.

EXAMPLE 113 Phosphoric acid, 2-bromoethyl 3-(hexadecylthio) propyl ester

A mixture of 2.0 g of 3-(hexadecylthio)-1-propanol, 2.7 g of2-bromoethyl phosphorodichloridate and 1.6 ml of triethylamine in 25 mlof carbon tetrachloride is stirred at room temperature for 3 hours.Following filtration through diatomaceous earth the solvents areevaporated and the residue stirred with 25 ml of 0.5M sodium acetate and25 ml of tetrahydrofuran at ambient temperature for 18 hours. Dilutehydrochloric acid is added followed by extraction with ethyl acetate.The combined extracts are dried and evaporated to give 3.0 g of thedesired product as a solid, m.p. 46°-49° C.

EXAMPLE 114 3,5-Dioxa-9-thia-4-phosphapentacosan-1-aminium,4-hydroxy-N,N,N-trimethyl-, hydroxide, inner salt, 4-oxide

A solution of 3.0 g of phosphoric acid, 2-bromoethyl3-(hexadecylthio)propyl ester in 36 ml of acetonitrile and 33 ml ofchloroform containing 31.4 ml of triethylamine is heated at 60° C. for18 hours. The solvents are evaporated and residue is stirred with 1.0 gof silver carbonate and 1.0 g of Amberlite®IR-4B in 35 ml of methylalcohol for 2 hours. The insolubles are filtered and the filtrateevaporated to a residue which is chromatographed on silica gel using70:30 chloroform:methyl alcohol and 65:35:6 chloroform:methyl alcohol:water to give upon evaporation of volatiles 1.5 g of the desired productas a white solid.

EXAMPLE 115 Ethyl 2-oxo-octadecanoate

A slurry of 40 ml of hexadecyl bromide in 250 ml of tetrahydrofurancontaining 3.64 g of magnesium chips is put in a sonic bath. Thissolution is added dropwise to a stirred solution of 35.3 ml ofdiethyloxalate in 70 ml of tetrahydrofuran at -15° C. over 1 hour. Thereaction mixture is poured into 3N hydrochloric acid and extracted withmethylene chloride. The extract is dried and evaporated. The concentrateis purified by chromatography on silica gel using 5% ethylacetate:petroleum ether. Fractions containing product are combined andevaporated. The residue is crystallized from petroleum ether to give21.9 g of the desired product as a solid.

EXAMPLE 116 Ethyl 2,2-difluorooctadecanoate

To a solution of 14.2 g of ethyl 2-oxo-octadecanoate in 100 ml ofmethylene chloride at 0° C. is added 20 g of diethylamino sulfurtrifluoride followed by stirring at ambient temperature for 72 hours.The mixture is partitioned between H₂ O and hexane. The hexane layer isdried and evaporated to an oil which is purified by chromatography onsilica gel using 0-25α ethylacetate in hexanes to give 6.9 g impureproduct and 7.9 g pure product as a waxy solid.

EXAMPLE 117 3.3-Difluoro-2-methyl-2-nonadecanol

To a solution of 6.9 g of ethyl 2,2-difluorooctadecanoate in 100 ml ofether at 0° C. is added 33 ml of methyl magnesium bromide dropwise withstirring. Following complete addition the reaction is allowed to reachroom temperature and stirred for 30 minutes. The reaction mixture iswashed with saturated ammonium chloride solution. The organic layer iswashed with 10% hydrochloric acid, water and 10% sodium bicarbonate. Theorganic layer is dried and evaporated to an oil which is purified bychromatography on silica gel using 1-10% ethyl acetate:hexanes to give3.8 g of the desired product as a solid following petroleum ethercrystallization.

EXAMPLE 118 3 3-Difluoro-2-methyl-1-nonadecene

A solution of 160 mg of 3,3-difluoro-2-methyl-2-nonadecanol in 2 ml ofpyridine containing 50 μl of phosphorous oxychloride is heated at 90° C.then allowed to cool to 50° C. An additional 250 μl of phosphorousoxychloride is added followed by heating at 90° C. for 2 hours. Afterallowing to stand at ambient temperature for 18 hours, the volatiles areremoved and the concentrate partitioned between hexanes and 10%hydrochloric acid, The organic layer is washed with water and 10% sodiumbicarbonate then dried and evaporated to a residue which is purified bychromatography on silica gel using hexane to give 140 mg of the desiredproduct as an oil.

EXAMPLE 119 2-(Bromomethyl)-3.3-difluoro-1-nonadecene

A slurry of 140 mg of 3,3-difluoro-2-methyl-1-nonadecene in 2 ml ofcarbon tetrachloride containing 43 mg of N-bromosuccinimide and acatalytic amount of azobisisobutryonitrile is stirred at ambienttemperature for 30 minutes. The mixture is filtered through a short padof silica gel which is repeatedly washed with hexane. The combinedfiltrates are evaporated and the residue is purified on silica gel usinghexanes to give 38 mg of the desired product as an oil.

EXAMPLE 120 Ethyl (3,3-difluoro-2-methylenenonadecyl)propanoate

To a slurry of 780 mg of hexane washed sodium hydride in 25 ml oftetrahydrofuran at 0° C. is added dropwise 2.2 ml of diethylmalonate.The cooling bath is removed and the reaction allowed to reach ambienttemperature. A solution of 1.9 g of 2-(bromomethyl)-3,3-difluoro-1-nonadecene in 10 ml of tetrahydrofuran is added dropwise over30 minutes followed by stirring for an additional 30 minutes. Ether isadded and the reaction washed with 10% hydrochloric acid, water andbrine. The organic layer is dried and evaporated to a gum which iscolumned on silica gel using 2% ethyl acetate:petroleum ether to give1.53 g of the desired product.

EXAMPLE 121 2-(3,3-Difluoro-2-methylenenonadecyl)-1,3-propanediol

To a slurry of 250 mg of lithium aluminum hydride in 10 ml of ether,cooled to 0° C. is added dropwise a solution of 1.48 g of ethyl(3,3-difluoro-2-methylene-nonadecyl)propanoate in 10 ml of etherfollowed by warming to ambient temperature, heating at reflux for 3hours, then allowing to cool to ambient temperature. The reactionmixture is washed with dilute hydrochloric acid, water and dilute sodiumbicarbonate, dried and evaporated to a gum which is purified on silicagel using 40-60% ethyl acetate:hexanes. The fractions containing productare collected, evaporated and the residue crystallized from hexanegiving 1.04 g of the desired product.

EXAMPLE 122 55.5-difluoro-2-(hexadecyloxy)methyl]-4-methylene-1-heneicosanol

To a slurry of 126 mg of hexane washed sodium hydride in 15 ml ofN,N-dimethylformamide is added 990 mg of2-(3,3-difluoro-2-methylenenonadecyl)-1,3propanediol with stirring. A0.96 ml portion of hexadecylbromide is added followed by heating at 80°C. for 2 hours, then cooled over an hour. Ammonium chloride is addedfollowed by evaporation of the solvent. The concentrate is dissolved inether and washed with 10% hydrochloric acid, water and brine. Theorganic layer is dried and evaporated to a gum which is purified onsilica gel using 1-100% ethyl acetate:hexanes to give 1.14 g of thedesired product as a gum.

EXAMPLE 123 3,5,9-Trioxa-4-phosphapentacosan-1-aminium,7-(3,3-difluoro-2-methylenenonadecyl)-4-hydroxy-N,N,N-trimethyl-,hydroxide, inner salt, 4-oxide

A solution of 1.10 g of5,5-difluoro-2-[(hexadecyloxy)methyl]-4-methylene-1-heneicosanol in 15ml of carbon tetrachloride, containing 0.42 ml of triethylamine and 0.37ml of 2-bromoethyl phosphorodichloridate is stirred at ambienttemperature for 3 hours. The solvent is evaporated and the residue isstirred for 18 hours at ambient temperature with 25 ml oftetrahydrofuran and 25 ml of 0.5M sodium acetate. The tetrahydrofuran isevaporated, ether added followed by washing with 10% hydrochloric acidand water. The organic layer is dried and evaporated to a gum. Added 15ml of chloroform and 40 ml of a solution of 50 g of trimethylamine in100 ml of acetonitrile followed by heating at 60° C. for 3 hours.Evaporated the volatiles and purified the concentrate on silica gelusing 3:7 methyl alcohol:chloroform followed by 5% water in 3:7 methylalcohol:chloroform. Volatiles are removed and the concentrate evaporatedwith toluene and dried to give 1.0 g of the desired product as a gum.

EXAMPLE 124 3,5,9-Trioxa-4-phosphapentacosan-1-aminium,7-(3,3-difluoro-2-oxononadecyl)-4-hydroxy-N,N,N-trimethyl-, hydroxide,inner salt, 4-oxide

A solution of 500 mg of 3,5,9-trioxa-4-phosphapentacosan-1-aminium,7-(3,3-difluoro-2-methylenenonadecyl)-4-hydroxy-N,N,N-trimethyl-,hydroxide, inner salt, 4-oxide in 20 ml of chloroform and 2 ml of ethylalcohol is cooled at -40° C., treated with ozone until blue and flushedwith oxygen. To the mixture is added 0.16 ml of trimethyl phosphitefollowed by evaporation. The concentrate is chromatographed on silicagel using 3:7 methyl alcohol:chloroform followed by 5% water in 3:7methyl alcohol:chloroform. Volatiles are removed to give 470 mg of thedesired product.

EXAMPLE 125 3,5.9-Trioxa-4-phosphapentacosan-1-aminium,7-(3,3-difluoro-2-hydroxynonadecyl)-4-hydroxy-N,N,N-trimethyl-,hydroxide, inner salt, 4-oxide

To a stirred 0° C. solution of 200 mg of3,5,9-trioxa-4-phosphapentacosan-1-aminium,7-(3,3-difluoro-2-oxononadecyl)-4-hydroxy-N,N,N-trimethyl-, hydroxide,inner salt, 4-oxide in 3.5 ml of chloroform is added dropwise 1.0 ml of1.0M borane-tetrahydrofuran complex followed by allowing to warm toambient temperature over 1 hour. The volatiles are removed, methylalcohol added and evaporated three times. The concentrate ischromatographed on silica gel with 3:7 methyl alcohol:chloroformfollowed by 5% water in 3:7 methyl alcohol:chloroform. Volatiles areremoved to give 194 mg of the desired product.

EXAMPLE 126 (S)-4-[(Hexadecyloxy)methyl]-2,2-dimethyl-1,3-dioxolane

To a stirred mixture 3.38 g of hexane washed sodium hydride in 67 ml ofdry toluene, under argon, is added dropwise 10.18 g of1,2-0-isopropylidene-sn-glycerol in 17 ml of toluene. After 15 minutes,a solution of 25.7 g of hexadecylbromide in 17 ml of toluene is addeddropwise over 20 minutes. The mixture is refluxed for 1.5 hours thenstirred at ambient temperature for 18 hours. Ice is added followed byice water. The organic layer was separated, washed with water, brine anddried. The solvent is removed to give 28 g of the desired product as apale yellow oil.

EXAMPLE 127 (R)-3-[hexadecyloxy)-1.2-propanediol

A stirred mixture of 28 g of(S)-4-[(hexadecyloxy)methyl]-2,2-dimethyl-1,3-dioxolane in 190 ml ofmethyl alcohol and 21 ml of 1N hydrochloric acid is refluxed for 2 hoursfollowed by cooling in an ice bath. The resulting solid is filtered andwashed with cold acetone. The solid is crystallized from acetone to give18.2 g of crystals which liquified at ambient temperature. The resultingoil is purified by chromatography on silica gel using 1:1 ethylacetate:hexanes. Fractions containing product are combined andevaporated to a material which is stirred with acetone at -20° C. togive 4.1 g of crystalline solid, m.p. 65°-66° C.

EXAMPLE 128 (S)-1-(hexadecyloxy)-3-[(4-methoxyphenyl)diphenylmethoxy]-2-propanol

To a solution of 4.1 g of (R)-3-(hexadecyloxy)-1,2-propanediol in 15 mlof dry pyridine is added dropwise with stirring under inert gas asolution of 5.6 g of p-methoxytrityl chloride in 15 ml oftetrahydrofuran. Stirring is continued for 1.5 hours at ambienttemperature then the reaction is stored in the refrigerator overnight.The reaction mixture is filtered and the cake washed with ether. Thefiltrate is evaporated to an oily residue which is dissolved inchloroform and washed with 10% sodium bicarbonate, water and then driedto give a stiff oil following evaporation of the solvent. The oil ispurified by high pressure liquid chromatography on silica gel using 1:9ethyl acetate:hexanes to give 7.6 g of the desired product as an oil.

EXAMPLE 129 Phosphoric acid, hexadecyl2-(hexadecyloxy)-1-[[(4-methoxyphenyl)diphenylmethoxy]1methyl]ethylmethyl ester

To a water bath cooled and stirred solution of 7.5 g of(S)-1-(hexadecyloxy)-3-[(4-methoxyphenyl)diphenylmethoxy]-2-propanol in30 ml of carbon tetrachloride is added, with stirring under inert gas,1.07 g of 4-dimethylaminopyridine and 7.63 ml of triethylamine. Asolution of 11.75 g of phosphorodichloridic acid, hexadecyl ester in 5ml of carbon tetrachloride is added at a rapid rate followed by washingthe addition funnel with 5 ml of carbon tetrachloride. The reaction isstirred at ambient temperature for 3 days. Added 9.1 ml of triethylamineto the reaction mixture and continued stirring for 2 hours. A 965 mlsolution of sodium bicarbonate (1 g/10 ml) is added to the reactionfollowed by three chloroform extractions. The combined extracts aredried and evaporated to give a dark oil which is purified by highpressure liquid chromatography on silica gel using 1:7 ethylacetate:hexanes. Fractions containing product are combined to give 8.2 gof the desired compound.

EXAMPLE 130

Phosphoric acid, hexadecyl 2-(hexadecyloxy)-1-(hydroxymethyl)ethylmethyl ester, (1R)-, upper diastereomer

Phosphoric acid, hexadecyl 2-(hexadecyloxy)-1-(hydroxymethyl-1ethylmethyl ester. (1R)- lower diastereomer

To a solution of 8.2 g of phosphoric acid, (S)-hexadecyl2-(hexadecyloxy)-1-[[(4-methoxyphenyl) diphenylmethoxy]methyl]ethylmethyl ester in 60 ml of methyl alcohol and 30 ml of chloroform, broughtto a boil, is added 4,9 g of Amberlist®-15 resin. The reaction isallowed to cool to ambient temperature while stirring over 90 minutes.The insolubles are filtered and the cake washed with 2:1 methylalcohol:chloroform. The combined filtrates are evaporated and theresidue dissolved in chloroform, washed with dilute sodium bicarbonateand dried. The volatiles are removed to give a yellow wax. The residueis purified by chromatography on silica gel using 1:1 ethylacetate:hexane, giving two major products. The upper fractions arecombined to give 2.31 g of phosphoric acid, hexadecyl2-(hexadecyloxy)-1-(hydroxy-methyl)ethyl methyl ester, (1R)-, upperdiasteromer of the desired product. The lower fractions are combined togive 2.06 g phosphoric acid, hexadecyl2-(hexadecyloxy)-1-(hydroxymethyl)ethyl methyl ester, (1R)-, lowerdiastereomer, of the desired product.

EXAMPLE 131 3,5,8,10-Tetraoxa-4 9-diphosphahexacosan-1-aminium,7-[(hexadecyloxy)methyl]-4-hydroxy-9-methoxy-N,N,N-trimethyl-,hydroxide, inner salt, 4,9-oxide, (7S)-, lower diastereomer

To a solution of 2.06 g of phosphoric acid, hexadecyl2-(hexadecyloxy)-1-(hydroxy-methyl)ethyl methyl ester, (1R)-, lowerdiastereomer in 35 ml of carbon tetrachloride stirred in an ice bathunder argon is added 1.02 g of 2-bromoethyl phosphorodichloridate in 2ml of carbon tetrachloride followed by the rapid addition of 0.63 ml oftriethylamine. The cooling bath is removed followed by stirring atambient temperature for 2 hours. The reaction is stored in therefrigerator overnight, followed by filtering through diatomaceousearth. The filter cake is washed well with toluene and the combinedfiltrates evaporated to a residue which is stirred at ambienttemperature with 80 ml of 0.5N sodium acetate and 80 ml oftetrahydrofuran for 2 hours. Evaporated the tetrahydrofuran andacidified the aqueous residue with hydrochloric acid then extracted withether several times. The combined extracts are washed with brine, driedand the solvent evaporated. The residue is evaporated with tolueneseveral times then dried under vacuum. The residue is dissolved in 75 mlof chloroform and 50 ml of a solution of 50 g of trimethylamine in 100ml of acetonitrile added followed by heating at 65° C. for 4acetonitrile added followed by heating at 65° C. for 4 hours. Thesolvents are removed and the residue dissolved in 38 ml of chloroformand 38 ml of methyl alcohol followed by adding 447 mg of silvercarbonate. The reaction mixture is stirred at ambient temperature for1.5 hours then filtered through diatomaceous earth. The cake is washedwith 1:1 chloroform:methyl alcohol. The combined filtrates areevaporated and the residue purified by chromatography on silica gelusing 30% methyl alcohol:chloroform and 65:35:6 chloroform:methylalcohol:water. Product fractions are combined and evaporated to aresidue which is stirred with ether and the resulting solid collected byfiltration to give 420 mg of the desired product as a white amorphouspowder.

EXAMPLE 132 3,5,8,10-Tetraoxa-4,9-diphosphahexacosan-1-aminium,7-(hexadecyloxy)methyl]-4-hydroxy-9-methoxy-N,N,N-trimethyl-, hydroxide,inner salt, 4,9-dioxide, (7S)-, upper diastereomer

A solution of 2.7 g of phosphoric acid, (1R)-hexadecyl2-(hexadecyloxy)-1-(hydroxymethyl)ethyl methyl ester, upper diastereomerin 38 ml of chloroform, 38 ml of methanol and 3.8 ml of water is stirredwith 502 mg of silver carbonate for 1.5 hours at ambient temperature.The mixture is filtered through diatomaceous earth and the cake washedwith 1:1 chloroform:methyl alcohol. The combined filtrates areevaporated and the residue purified by chromatography on silica gelusing 30% methyl alcohol:chloroform and 65:35:6 chloroform:methylalcohol:water. The concentrate of product fractions is stirred withether and the resulting solid collected by filtration to give 394 mg ofthe desired product as a white amorphous chloroform:methylalcohol:water. The concentrate of product fractions is stirred withether and the resulting solid collected by filtration to give 716 mg ofthe desired product as a white amorphous solid.

We claim:
 1. A compound, including the individual R and S enantiomer andthe racemic mixture, represented by the formula: ##STR31## wherein: A) nis an integer from 2 to 6;B) R is selected from the group consisting ofC₁ -C₄ alkyl; and C) Q is ##STR32## and W is hydrogen or hydroxyl, Z isCH₂ --OX wherein R¹ and X are selected from the group consisting of C₁-C₂₄ alkyl.
 2. A compound, including the individual R and S enantiomerand the racemic mixture, represented by the formula: ##STR33## wherein:A) n is an integer from 2 to 6;B) R is selected from the groupconsisting of C₁ -C₄ alkyl; and C) Z is --CH₂ --O--X wherein R¹ and Xare selected from the group consisting of C₁ -C₂₄ alkyl.
 3. Apharmaceutical composition of matter in unit dosage form comprising anamount of a compound according to claim 1 effective to inhibit thebiological effects of phospholipase A₂ or the mediators produced as aresult of its activity in association with a pharmaceutically acceptablecarrier.
 4. A pharmaceutical composition of matter in unit dosage formcomprising an amount of a compound according to claim 2 effective toinhibit the biological effects of phospohlipase A₂ or the mediatorsproduced as a result of its activity in association with apharmaceutically acceptable carrier.
 5. A method for inhibiting thebiological effects of phospholipase A₂ or the mediators produced as aresult of its activity which comprises administering to a mammal aneffective amount of a compound according to claim
 1. 6. A method forinhibiting the biological effects of phospholipase A₂ or the mediatorsproduced as a result of its activity which comprises administering to amammal an effective amount of a compound according to claim 2.